请使用支持JavaScript的浏览器! +,GeneSilencer® Transfection Reagent GeneSilencer® Transfection Reagent siRNA Transfection Reagent, siRNA Gene Silencing 蚂蚁淘商城
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genlantis/GeneSilencer/kit 0.18 ml/500020
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genlantis/GeneSilencer/kit 0.18 ml/500020
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genlantis
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500020
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4000-520-616
Description
Data
siRNA Design
Citations
  • High siRNA transfection efficiency
  • Functional gene silencing post siRNA delivery
  • Compatibility with diverse growth conditions (with and without serum)
  • Low cytotoxicity
  • Easy-to-use protocols for both adherent and suspension cells
GeneSilencer® siRNA Transfection Reagent is a novel cationic lipid formulation specifically designed for efficient delivery of siRNAs (small interfering RNAs) into a wide variety of cell types. siRNAs are short, gene-specific double-stranded RNAs that can cause gene silencing in mammalian cells by catalytically cleaving greater than 95% of the target mRNA (1,2,3).

Contents

  • Hydrated GeneSilencer Lipid
  • siRNA Diluent (corresponding ratio for each kit)

Storage

GeneSilencer siRNA Transfection Reagent is shipped at room temperature, as similar to other gene therapy products. For maximum stability, store all reagents at 4篊 upon receipt. If stored properly, all components are stable for 6 months.

GeneSilencer Transfection Reagent Kit

  • 50 rxn kit, 0.18 ml, T500020
  • 200 rxn kit, 0.75 ml, T500750
  • 1000 rxn kit, 5 x 0.75 ml, T505750
  • 10 rxn sample kit, 0.03 ml, T500020S

New Citations

Long Noncoding RNA pancEts-1 Promotes Neuroblastoma Progression through hnRNPK-Mediated β-Catenin Stabilization.Li, D.,et al (2018) Cancer Res. 78 (5): 1169-1183. Link

Apoptotic cells trigger the ABCA1/STAT6 pathway leading to PPAR‐γ expression and activation in macrophages.Kim, M-J.,et al (2018) J. Leukocyte Biol. 103(5): 885-895. Link

Transient Silencing of DNA Repair Genes Improves Targeted Gene Integration in the Filamentous Fungus Trichoderma reesei.Chum, P.Y.,et al ((2017) Appl. Environ. Microbiol. 83(15): e00535-17. Link

Programming of macrophages by apoptotic cancer cells inhibits cancer progression through exosomal PTEN and PPARγ ligands.Kim, Y-B.,et al (2017) bioRxiv 217562: 1-75. Link

Role of Sphingosine Kinase 1 and S1P Transporter Spns2 in HGF-mediated Lamellipodia Formation in Lung Endothelium.Fu, P.,et al (2016) J. Biol. Chem. 291(53): 27187?27203. Link

Ubiquitin-specific Protease 20 Regulates the Reciprocal Functions of β-Arrestin2 in Toll-like Receptor 4-promoted Nuclear Factor κB (NFκB) Activation.Jean-Charles, P-Y., (2016) J. Biol. Chem. 291 (14): 7450 ?7464. Link

Tolerogenic nanoparticles inhibit T cell杕ediated autoimmunity through SOCS2.Yeste, A.,et al (2016) Sci. Signal. 9(433): RA61. Link

Synthesis of an Endogenous Steroidal Na Pump Inhibitor Marinobufagenin, Implicated in Human Cardiovascular Diseases, Is Initiated by CYP27A1 via Bile Acid Pathway.Fedorova, O.V.,et al (2015) Circ Cardiovasc Genet. 8(5): 736-45. Link

Thioredoxin Activates MKK4-NFκB Pathway in a Redox-dependent Manner to Control Manganese Superoxide Dismutase Gene Expression in Endothelial Cells.Kundumani-Sridharan, V.,et al (2015) J. Biol. Chem. 290(28): 17505?17519. Link

Identification, Mechanism of Action, and Antitumor Activity of a Small Molecule Inhibitor of Hippo, TGF-β, and Wnt Signaling Pathways.Basu, D.,et al (2014) Mol. Cancer Ther. 13(6): 1457-1467. Link

PKC Potentiates Tyrosine Kinase Inhibitors STI571 and Dasatinib Cytotoxic Effect.Tob蚾, A.,et al (2014) Anticancer Res. 34(7): 3347-3356. Link

TREK2 Expressed Selectively in IB4-Binding C-Fiber Nociceptors Hyperpolarizes Their Membrane Potentials and Limits Spontaneous Pain.Acosta, C.,et al (2014) J. Neurosci. 34(4): 1494-1509. Link

RhoA/Phosphatidylinositol 3-Kinase/Protein Kinase B/Mitogen-Activated Protein Kinase Signaling after Growth Arrest朣pecific Protein 6/Mer Receptor Tyrosine Kinase Engagement Promotes Epithelial Cell Growth and Wound Repair via Upregulation of Hepatocyte Growth Factor in Macrophages.Lee, Y-J.,et al (2014) J. Pharma. Exp. Thera. 350(3): 563-577. Link

GeneSilencer Cell Line Citations: 3T3-L1, A549, A7, AGS, ASM Transformed, B16-F1, BEL7404, Bronchial C166, C2C12, C6R, CHO-K1, DC2.4, GS-KB-3-1, H22, H441, H460, H9c2, hCMEC/D3, HCP40, HCT-116, HEK 293, HEK 293T, HEK-293, HeLa, HepG2, HL-60, HMEC-1, HPAEC, HT29, HUT-78, JAR, Jurkat, KATOIII, L/Stab-2, LNCaP, M2, McA-RH 7777, MCF-7, MDA MB-231, MDA MB-468, MDA-MB-435, MG63, MKN 28, MKN 45, MV-4-11, Neuro2a, NIH-3T3, OK, PC-12, Cortical Neurons, Hippocampal Neurons, Macrophage, Pulmonary Artery, Endothelium, Dendritic Cells(Bone Marrow), Cortical Neurons, Coronary Artery Endothelial, Peritoneal Macrophage, Dorsal Root Ganglion Neurons, Retinal Ganglion, Dorsal Root Ganglion, Schwann Cells Cardiac Fibroblast, Cardiac Myocytes (Neonatal), Lung Microvascular Endothelial Cells, Aortic Muscle Cells, Dendritic Cells, Cerebellar Neurons, Osteoblast, HMVEC, Cardiac Myocytes, T-Cells (CD3+), Cortical Neurons, Dendritic Cells, Cortical Neurons, Aveolar Type II Cells, Macrophage, Dorsal Root Ganglion Lacrimal Gland Acinar Cells, Bone Marrow, Dendritic Cells, Pulmonary Artery Endothelium, Primary HPAEC. RAW 264.7, Renca, SaOs2, Saos-2, SCCH196, SNB19, SW480, U20S

GeneSilencerTM Reagent vs. siPORTTM NeoFXTM Reagent
HEK 293 Cells (click on images to enlarge)
HEK 293 Cells Only Negative Control
HEK 293 GeneSilencer+5nM GAPDHsiRNAHEK 293 siPORT NFX+5nM GAPDHsiRNA
HEK 293 GeneSilencer+10nM GAPDHsiRNAHEK 293 siPORT NFX+10nM GAPDHsiRNA
HEK 293 GeneSilencer+30nM GAPDHsiRNAHEK 293 siPORT NFX+30nM GAPDHsiRNA
HEK 293 GeneSilencer+50nM GAPDHsiRNAHEK 293 siPORT NFX+50nM GAPDHsiRNA

Figure Legend: HEK 293 cells were transfected with 5, 10, 30 or 50 nM of Silencer FAM Labeled GAPDH siRNA with either GeneSilencer (Genlantis) or siPORT NeoFX (Ambion) siRNA transfection reagents according to the manufacturer"s recommended protocols. Cells were incubated for 48 hours then fixed, permeablized, and incubated with AlexaFluor 555 phalloidin (Invitrogen), which stains cellular actin red. Cells were then mounted on slides and stained with DAPI (Invitrogen), which stains the cell nuclei blue. Transfected cells were visualized by fluorescence microscopy using identical exposure times for FITC, TRITC, and DAPI. The transfected GAPDH siRNA is localized in the cell cytosol and can be seen as green fluorescent specks or dots.

HeLa S3 Cells (click on images to enlarge)

HeLa GeneSilencer+5nMGAPDH siRNAHeLa siPORT 20NFX+5nM GAPDHsiRNA
HeLa GeneSilencer+10nM GAPDHsiRNA HeLa siPORT NFX+10nM GAPDHsiRNA
HeLa GeneSilencer+30nM GAPDHsiRNAHeLa siPORT NFX+30nM GAPDHsiRNA
HeLa GeneSilencer+50nM GAPDHsiRNAHeLa siPORT NFX+50nM GAPDHsiRNA

Figure Legend: HeLa S3 cells were transfected with 5, 10, 30 or 50 nM of Silencer FAM labeled GAPDH siRNA with either GeneSilencer (Genlantis) or siPORT&tm; NeoFX&tm; (Ambion) siRNA transfection reagents according to the manufacturer"s recommended protocols. Cells were incubated for 48 hours then fixed, permeablized, and incubated with AlexaFluor 555 phalloidin (Invitrogen), which stains cellular actin red. Cells were then mounted on slides and stained with DAPI (Invitrogen), which stains the cell nuclei blue. Transfected cells were visualized by fluorescence microscopy using identical exposure times for FITC, TRITC, and DAPI. The transfected GAPDH siRNA is localized in the cell cytosol and can be seen as green fluorescent specks or dots.

How to Design siRNA (Small Interfering RNA)

The sequence of the siRNA can be selected as follows:

  • 1. Start 75-100 bases downstream from the start codon of your gene of interest.
  • 2. Locate the first dimer.
  • 3. Record the next 19 nucleotides following the AA dimer.
  • 4. Calculate the percentage of G/C content of the AA-N19 21-base sequence. It must bebetween 30% and 70% with 50% being ideal. If the sequence does not meet the criteria,the search continues downstream to the next dimer until this condition is met.
  • 5. The 21-base sequence is subjected to a BLAST-search (NCBI database) against ESTlibraries of your organism to ensure that no other gene(s) is targeted. (The complement isautomatically searched as well.)
  • 6. If the conditions in either step 4 or 5 are not met, repeat steps 2 - 5.

    The sequence selection process has no other constraints. It is important to note that structure within thetargeted mRNA appears to have minimal effect on the availability of the mRNA target and efficacy of thesiRNA silencing approach. To-date, successful silencing has been achieved using the above method toselect the target sequence, although the method is essentially random with respect to accounting for mRNAstructure.Although siRNA silencing appears to be extremely effective by selecting a single target in the mRNA, itmay be desirable to design and employ two independent siRNA duplexes to control for specificity of thesilencing effect. This recommendation is only for specificity for it is yet unknown if the targeting of a geneby two different siRNA duplexes would be more effective than using a single siRNA duplex. It is believedthat the rate-limiting component of the siRNA effect is the availability of cellular nuclease components andnot mRNA target availability. Therefore, doubling the number of siRNA duplexes is not expected to doublethe rate or efficiency of silencing.If the selected siRNA duplex(es) do not function for silencing, the following steps are recommended. First,a search is conducted for sequencing errors in the gene and possible polymorphisms. Initial studies on thespecificity of target recognition by siRNA duplexes indicates that a single point mutation located in thepaired region of an siRNA duplex is sufficient to abolish target mRNA degradation. Second, a reexaminationis performed to confirm whether the cell line is from the expected species. Third, a secondand/or third target are selected and the corresponding siRNA duplexes prepared.

  • Cell Line Cell Type Source Genlantis Product Citation
    3T3-L1 Embryo Swiss Mouse GeneSilencer siRNA Transfection Reagent Mizuarai, S., Miki, S., Araki, H., Takahashi, K. and Kotani, H.(2005) Identification of Dicarboxylate Carrier Slc25a10 as Malate Transporter in de Novo Fatty Acid Synthesis. J. Biol. Chem.280(37): p. 32434-32441.
    A549 Lung Carcinoma Human GeneSilencer siRNA Transfection Reagent Ito, K., Yamamura, S., Essilfie-Quaye, S., Cosio, B., Ito, M., Barnes, P.J. and Adcock, I.M. (2006) Histone deacetylase 2-mediated deacetylation of the glucocorticoid receptor enables NFκB suppression. J. Exp. Med. 203(1): 7-13.
    A549 Lung Carcinoma Human GeneSilencer siRNA Transfection Reagent Stepulak, A., Sifringer, M., Rzeski, W., Endesfelder, S., Gratopp, A., Pohl, E.E., Bittigau, P., Hansen, H.H., Stryjecka-Zimmer, M., Turski, L. and Ikonomidou, C. (2005) NMDA antagonist inhibits the extracellular signal-regulated kinase pathway and suppresses cancer growth. Proc. Natl. Acad. Sci. 102: 15605 - 15610.
    A7 Melanoma Human GeneSilencer siRNA Transfection Reagent Scott, M.G.H., Pierotti, V., Storez, H., Lindberg, E., Thuret, A., Muntaner, O., Labbe-Jullie, C., Pitcher, J.A. and Marullo, S. (2006) Cooperative Regulation of Extracellular Signal-Regulated Kinase Activation and Cell Shape Change by Filamin A and β-Arrestins. Mol. & Cell Biol, 26: 3432-3445.
    AGS Gastric Epithelium Human GeneSilencer siRNA Transfection Reagent Nagasako, T., Sugiyama, T., Mizushima, T., Miura, Y., Kato, M. and Asaka, M. (2003) Up-regulated Smad5 Mediates Apoptosis of Gastric Epithelial Cells Induced by Helicobacter pylori Infection. J. Biol. Chem.: 278: 4821 - 4825.
    AGS Gastric Epithelium Human GeneSilencer siRNA Transfection Reagent Varro, A., Noble, P-J. M., Pritchard, D.M., Kennedy, S., Hart, C.A., Dimaline, R. and Dockray, G.J. (2004) Helicobacter pylori Induces Plasminogen Activator Inhibitor 2 in Gastric Epithelial Cells through Nuclear Factor-B and RhoA: Implications for Invasion and Apoptosis Cancer Res. 64: 1695 - 1702.
    ASM Transformed Airway Smooth Muscle Cells Human GeneSilencer siRNA Transfection Reagent Tran, T., Ens-Blackie, K., Rector, E.S., Stelmack, G.L., McNeill, K.D., Tarone, G., Gerthoffer, W.T., Unruh, H. and Halayko, A.J. (2007) Laminin-binding Integrin α7 is Required for Contractile Phenotype Expression by Human Airway Myocyte. Am. J. Respir. Cell Mol. Biol. 37(6): 668-80.
    B16-F1 Melanoma Mouse GeneSilencer siRNA Transfection Reagent Bertling, E., Hotulainen, P., Mattila, P.K., Matilainen, T., Salminen, M., and Lappalainen, P. (2004) Cyclase-associated-protein 1 (CAP1) promotes cofilin-induced actin dynamics in mammalian nonmuscle cells. Mol. Biol. Cell 15 (5): p. 2324-2334.
    B16-F1 Melanoma Mouse GeneSilencer siRNA Transfection Reagent Hotulainen, P., Paunola, E., Vartiainen, M.K. and Lappalainen, P. (2005) Actin-depolymerizing Factor and Cofilin-1 Play Overlapping Roles in Promoting Rapid F-Actin Depolymerization in Mammalian Nonmuscle Cells. Mol. Biol. Cell. 16(2): p. 649-664.
    BEL7404 Hepatoma Human GeneSilencer siRNA Transfection Reagent Liang, X-J, Finkel, T., Shen, D-W, Yin, J-J, Aszalos, A. and Gottesman, M.M. (2008) SIRT1 Contributes in Part to Cisplatin Resistance in Cancer Cells by Altering Mitochondrial Metabolism. Mol Cancer Res 6(9): 1499-506.
    Bronchial Smooth Muscle Human GeneSilencer siRNA Transfection Reagent Nunes, R.O., Schmidt, M., Dueck, G., Baarsma, H., Halayko, A.J., Kerstjens, H.A.M., Meurs, H. and Gosens, R. (2008) GSK-3/β-catenin signaling axis in airway smooth muscle: role in mitogenic signaling Am J Physiol Lung Cell Mol Physiol. 294: L1110 - L1118.
    C166 Yolk Sack Endothelium Mouse GeneSilencer siRNA Transfection Reagent Zhou, X., Stuart, A., Dettin, L.E., Rodriguez, G., Hoel, B. and Gallicano G.I. (2004) Desmoplakin is required for microvascular tube formation in culture. J. Cell Sci. 117: 3129-3140.
    C2C12 Myoblast Mouse GeneSilencer siRNA Transfection Reagent Evangelisti, C., Tazzari, P.L., Riccio, M., Fiume, R., Hozumi, Y., Fala, F., Goto, K.., Manzoli, L., Cocco, L. and Martelli, A.M. (2007) Nuclear diacylglycerol kinase-ζ is a negative regulator of cell cycle progression in C2C12 mouse myoblasts. FASEB J. 21: 3297 - 3307
    C6R Glioma Rat GeneSilencer siRNA Transfection Reagent Saarikangas, J., Hakanen, J., Mattila, P.K., Grumet, M., Salminen, M. and Lappalainen, P. (2008) ABBA regulates plasma-membrane and actin dynamics to promote radial glia extension. J Cell Sci. 121(Pt 9):1444-54.
    CHO-K1 Ovary Chinese Hamster GeneSilencer siRNA Transfection Reagent Kheifets, V., Bright, R., Inagaki, K., Schechtman, D. and Mochly-Rosen, D. (2006) Protein Kinase C δ PKC-Annexin V Interaction: a required step in δ PKC translocation and function. J. Biol. Chem. 281(32): p. 23218-23226.
    DC2.4 Bone Marrow Mouse GeneSilencer siRNA Transfection Reagent Jing, H., Yen, J-H and Ganea, D. (2004) A novel signaling pathway mediates the inhibition of CCL3/4 expression by PGE2. J. Biol. Chem. 279(53): p. 55176-55186.
    GS-KB-3-1 Epidermoid Adenocarcinoma Human GeneSilencer siRNA Transfection Reagent Liang, X-J, Finkel, T., Shen, D-W, Yin, J-J, Aszalos, A. and Gottesman, M.M. (2008) SIRT1 Contributes in Part to Cisplatin Resistance in Cancer Cells by Altering Mitochondrial Metabolism. Mol Cancer Res 6(9): 1499-506.
    H22 Hepatoma Mouse GeneSilencer siRNA Transfection Reagent Huang, B., Lei, Z., Zhang, G-M, Li, D., Song, C., Li, B., Liu, Y., Yuan, Y., Unkeless, J., Xiong, H. and Feng, Z-H (2008) SCF-mediated mast cell infiltration and activation exacerbate the inflammation and immunosuppression in tumor microenvironment. Blood 112(4): 1269-79.
    H22 Hepatoma Mouse GeneSilencer siRNA Transfection Reagent Huang, B., Zhao, J., Shen, S., Li, H., He, K-L, Shen, G-X, Mayer, L., Unkeless, J., Li, D., Yuan, Y., Zhang, G.-M., Xiong, H. and Feng, Z.-H. (2007) Listeria monocytogenes Promotes Tumor Growth via Tumor Cell Toll-Like Receptor 2 Signaling. Cancer Res. 67(9): p. 4346-4352.
    H441 Lung Adenocarcinoma Human GeneSilencer siRNA Transfection Reagent Zhang, Y-A., Nemunaitis, J., Samuel, S.K., Chen, P., Shen, Y. and Tong, A.W. (2006) Antitumor Activity of an Oncolytic Adenovirus-Delivered Oncogene Small Interfering RNA. Cancer Res. 66(19): 9736-9743.
    H460 Lung Cancer Human GeneSilencer siRNA Transfection Reagent Ren, J., Shi, M., Liu, R., Yang, Q-H., Johnson, T., Skarnes, W.C. and Du, C. (2005) The Birc6 (Bruce) gene regulates p53 and the mitochondrial pathway of apoptosis and is essential for mouse embryonic development. Proc. Natl. Acad. Sci. USA. 102(3): p. 565-570.
    H9c2 Myoblast Rat GeneSilencer siRNA Transfection Reagent Saotome, M., Safiulina, D., Szabadkai, G., Das, S., Fransson, A., Aspenstrom, P., Rizzuto, R. and Hajnoczky, G. (2008) Bidirectional Ca2+-dependent control of mitochondrial dynamics by the Miro GTPase. Proc Natl Acad Sci U S A 105(52): p. 20728.
    hCMEC/D3 Brain Microvascular Endothelia Human GeneSilencer siRNA Transfection Reagent Huang, W., Eum, S.Y., Andras, I.E., Hennig, B. and Toborek, M. (2009) PPAR{alpha} and PPAR{gamma} attenuate HIV-induced dysregulation of tight junction proteins by modulations of matrix metalloproteinase and proteasome activities. FASEB J 13 Jan 2009.
    HCP40 Colon adenocarcinoma null GeneSilencer siRNA Transfection Reagent Yao, K., Shida, S., Selvakumaran, M., Zimmerman, R., Simon, E., Schick, J., Haas, N.B., Balke, M., Ross, H., Johnson, S.W. and O"Dwyer, P.J. (2005) Macrophage Migration Inhibitory Factor Is a Determinant of Hypoxia-Induced Apoptosis in Colon Cancer Cell Lines. Clin Cancer Res. 11 (20): 7264 - 7272.
    HCT-116 Colon Carcinoma Human GeneSilencer siRNA Transfection Reagent Wang, J., Rajput, A., Kan, J.L.C., Rose, R., Liu, X., Kuropatwinski, K., Hauser, J., Beko, A., Dominquez, I., Sharratt, E.A., Brattain, L., LeVea, C., Sun, F., Keane, D.M., Gibson, N.W. and Michael G. Brattain (2009) Knockdown of Ron kinase inhibits mutant PI3 kinase and reduces metastasis in human colon carcinoma. J. Biol. Chem. 284 (16) 10912-10922.
    HCT-116 Colon Adenocarcinoma Human GeneSilencer siRNA Transfection Reagent Yao, K., Shida, S., Selvakumaran, M., Zimmerman, R., Simon, E., Schick, J., Haas, N.B. Balke, M., Ross, H., Johnson, S.W. and O"Dwyer, P.J. (2005) Macrophage Migration Inhibitory Factor Is a Determinant of Hypoxia-Induced Apoptosis in Colon Cancer Cell Lines. Clin. Can. Res. 11: 7264-7272.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Ahn, S., Nelson, C.D., Garrison, T.R., Miller, W.E. and Lefkowitz, R.J. (2003) Desensitization, internalization, and signaling functions of -arrestins demonstrated by RNA interference. Proc. Natl. Acad. Sci. 100: 1740 - 1744.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Ahn, S., Shenoy, S.K., Wei, H. and Lefkowitz, R.J. (2004) Differential Kinetic and Spatial Patterns of {beta}-Arrestin and G Protein-mediated ERK Activation by the Angiotensin II Receptor. J. Biol. Chem. 279 (34): 35518-35525.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Ahn, S., Wei, H., Garrison, T.R. and Lefkowitz, R.J. (2004) Reciprocal Regulation of Angiotensin Receptor-activated Extracellular Signal-regulated Kinases by (beta)-Arrestins 1 and 2. J. Biol. Chem. 279: 7807-7811.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Khundmiri, S.J., Dean, W.L., McLeish, K.R. and Lederer, E.D. (2005) Parathyroid Hormone-mediated Regulation of Na+-K+-ATPase Requires ERK-dependent Translocation of Protein Kinase C{alpha}. J. Biol. Chem. 280(10): p. 8705-8713.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Kim, J., Ahn, S., Ren, X-R., Whalen, E.J., Reiter, E., Wei, H. and Lefkowitz, R.J. (2005) Functional antagonism of different G protein-coupled receptor kinases for {beta}-arrestin-mediated angiotensin II receptor signaling. Proc. Natl. Acad. Sci.驴102: 1442 - 1447.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Kohout, T.A., Nicholas, S.L., Perry, S.J., Reinhart, G., Junger, S. and Struthers, R. (2004) Differential Desensitization, Receptor Phosphorylation, {beta}-Arrestin Recruitment, and ERK1/2 Activation by the Two Endogenous Ligands for the CC Chemokine Receptor 7. J. Biol. Chem. 279 (22): p. 23214-23222.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Liang, Y., Yu, W., Li, Y., Yan, X., Huang, Q. and Zhu, X. (2004) Nudel functions in membrane traffic mainly through association with Lis1 and cytoplasmic dynein. J. Cell Biol. 164 (4): 557-566.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Ren, X-R., Reiter, E., Ahn, S., Kim, J., Chen, W. and Lefkowitz, R.J. (2005) Different G protein-coupled receptor kinases govern G protein and {beta}-arrestin-mediated signaling of V2 vasopressin receptor. Proc. Natl. Acad. Sci. 102: 1448 - 1453.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Shenoy, S.K., Drake, M.T., Nelson, C.D., Houtz, D.A., Xiao, K., Madabushi, S., Reiter, E., Premont, R.T., Lichtarge, O. and Lefkowitz, R.J. (2006) {beta}-Arrestin-dependent, G Protein-independent ERK1/2 Activation by the {beta}2 Adrenergic Receptor. J. Biol. Chem. 281(2): 1261-1273.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Shikama, Y., Yamada, M., Miyashita, T. (2004) Caspase-8 and caspase-10 activate NF-B through RIP, NIK and IKK kinases. Eur. J. Immun. 33 (7): 1998-2006.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Wei, H., Ahn, S., Barnes, W.G. and Lefkowitz, R.J., (2004) Stable interaction between beta -arrestin2 and AT 1A receptor is required for beta -arrestin2 mediated activation of extracellular signal-regulated kinase 1 and 2. J. Biol. Chem. 279: 48255-48261.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Wei, H., Ahn, S., Shenoy, S.K., Karnik, S.S., Hunyady, L., Luttrell, L.M. and Lefkowitz, R.J. (2003) Independent -arrestin 2 and G protein-mediated pathways for angiotensin II activation of extracellular signal-regulated kinases 1 and 2. Proc. Natl. Acad. Sci. 100: 10782 - 10787.
    HEK 293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Wu, J-H, Goswami, R., Kim, L.K., Miller, W.E., Peppel, K. and Freedman, N.J. (2005) The Platelet-derived Growth Factor Receptor-{beta} Phosphorylates and Activates G Protein-coupled Receptor Kinase-2: a Mechanism for Feedback Inhibition. J. Biol. Chem. 280(35): p. 31027-31035.
    HEK 293T Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Wei, H., Ahn, S., Shenoy, S.K., Karnik, S.S., Hunyady, L., Luttrell, L.M. and Lefkowitz, R.J. (2003) Independent -arrestin 2 and G protein-mediated pathways for angiotensin II activation of extracellular signal-regulated kinases 1 and 2. Proc. Natl. Acad. Sci. 100: 10782 - 10787.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Ahn, S., Kim, J., Hara, M.R., Ren, X. and Lefkowitz, R.J. (2009) β -arrestin2 mediates anti-apoptotic signaling through regulation of bad phosphorylation. J Biol Chem 284(13): 8855-65.
    HEK-293 Embryonic Fibroblast Human GeneSilencer siRNA Transfection Reagent Barthet, G., Framery, B., Gaven, F., Pellissier, L., Reiter, E., Claeysen, S., Bockaert, J. and Dumuis, A. (2007) 5-Hydroxytryptamine 4 Receptor Activation of the Extracellular Signal-regulated Kinase Pathway Depends on Src Activation but Not on G Protein or beta-Arrestin Signaling. Mol. Biol. Cell. 18(6): 1979-1991.
    HEk-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent El-Shewy, H.M., Johnson, K.R., Lee, M-H, Jaffa, A.A., Obeid, L.M. and Luttrell, L.M. (2006) Insulin-like Growth Factors Mediate Heterotrimeric G Protein-dependent ERK1/2 Activation by Transactivating Sphingosine 1-Phosphate Receptors . J. Biol. Chem. 281(42): 31399-31407.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent El-Shewy, H.M., Lee, M-H., Obeid, L.M., Jaffa, A.A. and Luttrell, L.M. (2007) The Insulin-like Growth Factor Type 1 and Insulin-like Growth Factor Type 2/Mannose-6-phosphate Receptors Independently Regulate ERK1/2 Activity in HEK293 Cells. J. Biol. Chem. 282(36): 26150-26157.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Gesty-Palmer, D., Chen, M., Reiter, E., Ahn, S., Nelson, C.D., Wang, S., Eckhardt, A.E., Cowan, C.L., Spurney, R.F., Luttrell, L.M. and Lefkowitz, R.J. (2006) Distinct beta-Arrestin- and G Protein-dependent Pathways for Parathyroid Hormone Receptor-stimulated ERK1/2 Activation. J. Biol. Chem. 281(16): 10856-10864.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent IM Kim, DG Tilley, J Chen, NC Salazar, EJ Whalen, JD Violin, and HA Rockman (2008) Beta-blockers alprenolol and carvedilol stimulate beta-arrestin-mediated EGFR transactivation. Proc Natl Acad Sci 105(38): 14555-60.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Kani, S., Nakayama, E., Yoda, A., Onishi, N., Sougawa, N., Hazaka, Y., Umeda, T., Takeda, K., Ichijo, H., Hamada, Y. and Minami, Y. (2007) Chk2 kinase is required for methylglyoxal-induced G2/M cell-cycle checkpoint arrest: implication of cell-cycle checkpoint regulation in diabetic oxidative stress signaling. Genes Cells. 12(8): 919-928.
    HEk-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Kara, E., Crepieux, P., Gauthier, C., Martinat, N., Piketty, V., Guillou, F., and Reiter, E. (2006) A Phosphorylation Cluster of Five Serine and Threonine Residues in the C-Terminus of the Follicle-Stimulating Hormone Receptor Is Important for Desensitization But Not for β-Arrestin-Mediated ERK Activation. Mol. Endocrinol. 20(11): p. 3014-3026.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Nelson, C.D., Kovacs, J.J., Nobles, K.N., Whalen, E.J. and Lefkowitz, R.J. (2008) β-Arrestin Scaffolding of Phosphatidylinositol 4-Phosphate 5-Kinase Iα Promotes Agonist-stimulated Sequestration of the {beta}2-Adrenergic Receptor. J. Biol. Chem. 283(30): p. 21093-21101.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Noma, T., Lemaire, A., Prasad, S.V.N., Barki-Harrington, L., Tilley, D.G., Chen, J., Le Corvoisier, P., Violin, J.D., Wei, H., Lefkowitz, R.J. and Rockman, H.A. (2007) β-Arrestin mediated β1-adrenergic receptor transactivation of the EGFR confers cardioprotection. J. Clin. Invest. 117(9): p. 2445-2458.
    HEk-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Rajagopal, K., Whalen, E.J., Violin, J.D., Stiber, J.A., Rosenberg, P.B., Premont, R.T., Coffman, T.M., Rockman, H.A. and Lefkowitz, R.J. (2006) β-Arrestin2-mediated inotropic effects of the angiotensin II type 1A receptor in isolated cardiac myocytes. PNAS. 103: 16284 - 16289.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Shenoy, S.K., Xiao, K., Venkataramanan, V., Snyder, P.M., Freedman, N.J. and Weissman, A.M. (2008) NEDD4 mediates agonist-dependent ubiquitination, lysosomal targeting and degradation of the beta 2 adrenergic receptor. J. Biol. Chem. 283: 22166 - 22176.
    HEK-293 Embryonic Kidney Human GeneSilencer siRNA Transfection Reagent Violin, J.D., DeWire, S.M., Barnes, W.G. and Lefkowitz, R.J. (2006) G Protein-Coupled Receptor Kinase and β-arrestin mediated desensitization of the angiotensin II type 1A receptor elucidated by diacylgylcerol dynamics. J. Biol. Chem. 281: 36411 - 36419.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Bentley, A.M., Normand, G., Hoyt, J. and King, R.W. (2007) Distinct Sequence Elements of Cyclin B1 Promote Localization to Chromatin, Centrosomes, and Kinetochores during Mitosis. Mol. Biol. Cell, Dec 2007; 18: 4847 - 4858.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Cassimeris, L. and Morabito, J. (2004) TOGp, the Human Homolog of XMAP215/Dis1, Is Required for Centrosome Integrity, Spindle Pole Organization, and Bipolar Spindle Assembly. Mol. Biol. Cell 15 (4): 1580-1590.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Chung, J-S., Sato, K., Dougherty, I.I., Cruz, Jr., P.D. and Ariizumi, K. (2007) DC-HIL is a negative regulator of T lymphocyte activation. Blood. 109(10): p. 4320-4327.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Kheifets, V., Bright, R., Inagaki, K., Schechtman, D. and Mochly-Rosen, D. (2006) Protein Kinase C δ}PKC)-Annexin V Interaction: a required step in δ PKC translocation and function. J. Biol. Chem. 281(32): p. 23218-23226.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Layzer, J.M., McCaffrey, A.P., Tanner, A.K., Huang, Zan, Kay, Mark A. and Sullenger, B.A. (2004) In vivo activity of nuclease-resistant siRNAs. RNA 10 (5): p. 766-771.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Pomerening, J.R., Ubersax, J.A. and Ferrell, J.E., Jr. (2008) Rapid Cycling and Precocious Termination of G1 Phase in Cells Expressing CDK1AF. Mol. Biol. Cell, 19: 3426 - 3441.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Shulga, N. and Pastorino, J.G. (2006) Acyl Coenzyme A-binding Protein Augments Bid-induced Mitochondrial Damage and Cell Death by Activating μ-Calpain. J. Biol. Chem. 281(41): 30824-30833.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Witherow, D.S., Garrison, T.R., Miller, W.E. and Robert J. Lefkowitz (2004) {beta}-Arrestin inhibits NF-{kappa}B activity by means of its interaction with the NF-{kappa}B inhibitor I{kappa}B{alpha}. Proc. Natl. Acad. Sci. USA 101: 8603-8607.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Yang, Q-H, Church-Hajduk, R., Ren, J., Newton, M.L. and Du, C. (2003) Omi/HtrA2 catalytic cleavage of inhibitor of apoptosis (IAP) irreversibly inactivates IAPs and facilitates caspase activity in apoptosis. Genes & Dev. 17: 1487 - 1496.
    HeLa Cervical Carcinoma Human GeneSilencer siRNA Transfection Reagent Zhu, S., Wang, W., Clarke, D.C. and Liu, X. (2007) Activation of Mps1 Promotes Transforming Growth Factor-β-independent Smad Signaling. J. Biol Chem. 282 (25) 18327-18338.
    HepG2 Hepatocellular Carcinoma Human GeneSilencer siRNA Transfection Reagent Pastorino, J.G. and Shulga, N. (2008) TNFalpha can provoke cleavage and activation of sterol regulatory element binding protein in ethanol exposed cells via a caspase dependent pathway that is cholesterol-insensitive. J. Biol. Chem. 283: 25638 - 25649.
    HL-60 Peripheral Blood Promyelocytic Leukemia Human GeneSilencer siRNA Transfection Reagent Smirnova, I.V., Kajstura, M., Sawamura, T. and Goligorsky, M.S. (2004) Asymmetric dimethylarginine upregulates LOX-1 in activated macrophages: role in foam cell formation. AJP: Heart 287(2): p. H782-H790
    HMEC-1 Human Microvascular Endothelial Cells Human GeneSilencer siRNA Transfection Reagent Zhou, X., Stuart, A., Dettin, L.E., Rodriguez, G., Hoel, B. and Gallicano G.I. (2004) Desmoplakin is required for microvascular tube formation in culture. J. Cell Sci. 117: 3129-3140.
    HPAEC Pulmonary Artery Endothelium Human GeneSilencer siRNA Transfection Reagent Berdyshev, E.V., Gorshkova, I., Skobeleva, A., Bittman, R., Lu, X., Dudek, S.M., Mirzapoiazova, T., Garcia, J.G.N. and Natarajan, V. (2009) FTY720 inhibits ceramide synthases and upregulates dihydrosphingosine-1-phosphate formation in human lung endothelial cells. J. Biol. Chem. 284 (9): 5467-5477.
    HT29 Colon Adenocarcinoma Human GeneSilencer siRNA Transfection Reagent Yao, K., Shida, S., Selvakumaran, M., Zimmerman, R., Simon, E., Schick, J., Haas, N.B. Balke, M., Ross, H., Johnson, S.W. and O"Dwyer, P.J. (2005) Macrophage Migration Inhibitory Factor Is a Determinant of Hypoxia-Induced Apoptosis in Colon Cancer Cell Lines. Clin. Can. Res. 11: 7264-7272.
    HUT-78 T Cells Human GeneSilencer siRNA Transfection Reagent Maneechotesuwan, K., Xin, Y., Ito, K., Jazrawi, E., Lee, K-Y., Usmani, O.S., Barnes, P.J. and Adcock, I.M. (2007) Regulation of Th2 Cytokine Genes by p38 MAPK-Mediated Phosphorylation of GATA-3. J. Immunol. 178(4): 2491-2498.
    JAR Choriocarcinoma Human GeneSilencer siRNA Transfection Reagent Abboud-Jarrous, G., Atzmon, R., Peretz, T., Palermo, C., Gadea, B.B., Joyce, J.A. and Vlodavsky, I. (2008) Cathepsin L Is Responsible for Processing and Activation of Proheparanase through Multiple Cleavages of a Linker Segment. J. Biol. Chem. 283: 18167 - 18176.
    Jurkat T-Cell Lymphoma Human GeneSilencer siRNA Transfection Reagent Nguyen, J.T. and Wells, J.A. (2003) Direct activation of the apoptosis machinery as a mechanism to target cancer cells. Proc. Natl. Acad. Sci. 100: 7533  7538.
    KATOIII Gastric Epithelium Human GeneSilencer siRNA Transfection Reagent Nagasako, T., Sugiyama, T., Mizushima, T., Miura, Y., Kato, M. and Asaka, M. (2003) Up-regulated Smad5 Mediates Apoptosis of Gastric Epithelial Cells Induced by Helicobacter pylori Infection. J. Biol. Chem: 278: 4821 - 4825.
    L/Stab-2 Fibroblast Mouse GeneSilencer siRNA Transfection Reagent Park, S-Y., Kang, K-B., Thapa, N., Kim, S-Y., Lee, S-J. and Kim, I-S. (2008) Requirement of Adaptor Protein GULP during Stabilin-2-mediated Cell Corpse Engulfment. J. Biol. Chem. 283(16): 10593-10600.
    LNCaP Prostate Cancer Human GeneSilencer siRNA Transfection Reagent Takayama, K., Tsutsumi, S., Suzuki, T., Horie-Inoue, K., Ikeda, K., Kaneshiro, K. Fujimura, T., Kumagai, J., Urano, T., Sakaki, Y., Shirahige, K., Sasano, H., Takahashi, S., Kitamura, T., Ouchi, Y., Aburatani, H. and Inoue, S. (2009) Amyloid precursor protein is a primary androgen target gene that promotes prostate cancer growth. Cancer Res 1 69(1): p. 137.
    M2 Melanoma Human GeneSilencer siRNA Transfection Reagent Scott, M.G.H., Pierotti, V., Storez, H., Lindberg, E., Thuret, A., Muntaner, O., Labbe-Jullie, C., Pitcher, J.A. and Marullo, S. (2006) Cooperative Regulation of Extracellular Signal-Regulated Kinase Activation and Cell Shape Change by Filamin A and β-Arrestins. Mol. & Cell Biol, 26: 3432-3445.
    McA-RH 7777 Hepatoma Rat GeneSilencer siRNA Transfection Reagent Pastorino, J.G. and Shulga, N. (2008) Tumor Necrosis Factor-{alpha} Can Provoke Cleavage and Activation of Sterol Regulatory Element-binding Protein in Ethanol-exposed Cells via a Caspase-dependent Pathway That Is Cholesterol Insensitive. J. Biol. Chem. 283: 25638 - 25649.
    MCF-7 Breast Adenocarcinoma Human GeneSilencer siRNA Transfection Reagent Ikeda, K., Ogawa, S., Tsukui, T., Horie-Inoue, K., Ouchi, Y., Kato, S., Muramatsu, M. and Inoue, S. (2004) Protein Phosphatase 5 Is a Negative Regulator of Estrogen Receptor-mediated Transcription. Mol. Endocrinol. 18(5): 1131-1143.
    MDA MB-231 Breast Cancer Human GeneSilencer siRNA Transfection Reagent Ge, L., Shenoy, S.K., Lefkowitz, R.J. and DeFea, K.A. (2004) Constitutive protease-activated-receptor-2 mediated migration of MDA MB-231 breast cancer cells requires both beta-arrestin-1 and 2. J. Biol. Chem. 279(53): p. 55419-55424.
    MDA MB-468 Breast Carcinoma Human GeneSilencer siRNA Transfection Reagent Ge, L., Shenoy, S.K., Lefkowitz, R.J. and DeFea, K.A. (2004) Constitutive protease-activated-receptor-2 mediated migration of MDA MB-231 breast cancer cells requires both beta-arrestin-1 and 2. J. Biol. Chem.279(53): p. 55419-55424.
    MDA-MB-435 Melanoma Human GeneSilencer siRNA Transfection Reagent Abboud-Jarrous, G., Atzmon, R., Peretz, T., Palermo, C., Gadea, B.B., Joyce, J.A.and Vlodavsky, I. (2008) Cathepsin L Is Responsible for Processing and Activation of Proheparanase through Multiple Cleavages of a Linker Segment. J. Biol. Chem. 283: 18167 - 18176.
    MDA-MB-468 Breast Carcinoma Human GeneSilencer siRNA Transfection Reagent Zoudilova, M., Kumar, P., Ge, L., Wang, P., Bokoch, G. M. and DeFea, K. A. (2007) β-arrestin-dependent regulation of the cofilin pathway downstream of protease-activated receptor-2. J. Biol. Chem. 282: 20634 - 20646.
    MG63 Osteosarcoma Human GeneSilencer siRNA Transfection Reagent Ichikawa, T., Horie-Inoue, K., Ikeda, K., Blumberg, B. and Inoue, S. (2007) Vitamin K2 induces phosphorylation of protein kinase A and expression of novel target genes in osteoblastic cells. J Mol Endocrinol 1 39 (4): 239.
    MKN 28 Gastric Epithelium Human GeneSilencer siRNA Transfection Reagent Nagasako, T., Sugiyama, T., Mizushima, T., Miura, Y., Kato, M. and Asaka, M. (2003) Up-regulated Smad5 Mediates Apoptosis of Gastric Epithelial Cells Induced by Helicobacter pylori Infection. J. Biol. Chem.: 278: 4821 - 4825.
    MKN 45 Gastric Epithelium Human GeneSilencer siRNA Transfection Reagent Nagasako, T., Sugiyama, T., Mizushima, T., Miura, Y., Kato, M. and Asaka, M. (2003) Up-regulated Smad5 Mediates Apoptosis of Gastric Epithelial Cells Induced by Helicobacter pylori Infection. J. Biol. Chem.: 278: 4821 - 4825.
    MV-4-11 Biophenotypic B Myelomonocytic Leukemia Human GeneSilencer siRNA Transfection Reagent Yang, X., Liu, L., Sternberg, D., Tang, L., Galinsky, I., DeAngelo, D. and Stone, R. (2005) The FLT3 Internal Tandem Duplication Mutation Prevents Apoptosis in Interleukin-3-Deprived BaF3 Cells Due to Protein Kinase A and Ribosomal S6 Kinase 1-Mediated BAD Phosphorylation at Serine 112. Cancer Res. 65(16): p. 7338-7347.
    Neuro2a Neuroblastoma Mouse GeneSilencer siRNA Transfection Reagent Bertling, E., Hotulainen, P., Mattila, P.K., Matilainen, T., Salminen, M., and Lappalainen, P. (2004) Cyclase-associated-protein 1 (CAP1) promotes cofilin-induced actin dynamics in mammalian nonmuscle cells. Mol. Biol. Cell 15 (5): p. 2324-2334.
    NIH-3T3 Fibroblast Mouse GeneSilencer siRNA Transfection Reagent Bertling, E., Hotulainen, P., Mattila, P.K., Matilainen, T., Salminen, M., and Lappalainen, P. (2004) Cyclase-associated-protein 1 (CAP1) promotes cofilin-induced actin dynamics in mammalian nonmuscle cells. Mol. Biol. Cell 15 (5): 2324-2334.
    NIH-3T3 Fibroblast Mouse GeneSilencer siRNA Transfection Reagent Hotulainen, P., Paunola, E., Vartiainen, M.K. and Lappalainen, P. (2005) Actin-depolymerizing Factor and Cofilin-1 Play Overlapping Roles in Promoting Rapid F-Actin Depolymerization in Mammalian Nonmuscle Cells. Mol. Biol. Cell. 16(2): p. 649-664.
    NIH-3T3 Fibroblast Mouse GeneSilencer siRNA Transfection Reagent Nomachi, A., Nishita, M., Inaba, D., Enomoto, M., Hamasaki, M. and Minami, Y. (2008) Receptor Tyrosine Kinase Ror2 Mediates Wnt5a-induced Polarized Cell Migration by Activating c-Jun N-terminal Kinase via Actin-binding Protein Filamin A. J. Biol. Chem. 283(41): 27973-27981.
    OK Kidney Opossum GeneSilencer siRNA Transfection Reagent Khundmiri, S.J., Ameen, M., Delamere, N.A. and Lederer, E.D. (2008) PTH-mediated regulation of Na+ -K+ -ATPase requires Src kinase-dependent ERK Phosphorylation. Am J Physiol Renal Physiol. 295(2): F426-F437.
    OK Kidney Opossum GeneSilencer siRNA Transfection Reagent Khundmiri, S.J., Dean, W.L., McLeish, K.R. and Lederer, E.D. (2005) Parathyroid Hormone-mediated Regulation of Na+-K+-ATPase Requires ERK-dependent Translocation of Protein Kinase C{alpha}. J. Biol. Chem. 280(10): p. 8705-8713.
    PC-12 Pheochromocytoma Rat GeneSilencer siRNA Transfection Reagent de Barry, J., Janoshazi, A., Luc Dupont, J., Procksch, O., Chasserot-Golaz, S., Jeromin, A. and Vitale, N. (2006) Functional Implication of Neuronal Calcium Sensor-1 and Phosphoinositol 4-Kinase-β Interaction in Regulated Exocytosis of PC12 Cells. J. Biol Chem. 281(26): 18098-18111.
    Primary Cortical Neurons Mouse GeneSilencer siRNA Transfection Reagent Aarts, M., Lihara, K., Wei, W-L, Xiong, Z-G, Arundine, M., Cerwinski, W., MacDonald, J.F., Tymianski, M. (2003) A Key Role for TRPM7 Channels in Anoxic Neuronal Death. Cell 115: 863.
    Primary Cortical Neurons Mouse GeneSilencer siRNA Transfection Reagent Aleyasin, H., Cregan, S.P., Iyirhiaro, G., OHare, M.J., Callaghan, S.M., Slack, R.S. and Park, D.S. (2004) Nuclear Factor-B Modulates the p53 Response in Neurons Exposed to DNA Damage. J. Neurosci. 24 (12): 2963-2973.
    Primary Hippocampal Neurons Rat GeneSilencer siRNA Transfection Reagent Amaral, M.D. and Pozzo-Miller, L. (2007) TRPC3 Channels Are Necessary for Brain-Derived Neurotrophic Factor to Activate a Nonselective Cationic Current and to Induce Dendritic Spine Formation. J. Neurosci. 27(19): p. 5179-5189.
    Primary Macrophage Human GeneSilencer siRNA Transfection Reagent Asmis, R., Wang, Y., Xu, L., Kisgati, M., Begley, J.G. and驴Mieyal, J.J. (2005) A novel thiol oxidation-based mechanism for adriamycin-induced cell injury in human macrophages. FASEB J. published 13 September 2005, 10.1096/fj.04-2991fje.
    Primary Pulmonary Artery Endothelium Human GeneSilencer siRNA Transfection Reagent Birukova, A.A., Chatchavalvanich, S., Rios, A., Kawkitinarong, K., Garcia, J.G.N. and Birukov, K.G. (2006) Differential Regulation of Pulmonary Endothelial Monolayer Integrity by Varying Degrees of Cyclic Stretch. Am. J. Pathol. 168: 1749 - 1761.
    Primary Cortical Neurons Mouse GeneSilencer siRNA Transfection Reagent Burkhalter, J., Fiumelli, H., Erickson, J.D. and Martin, J-L. (2007) A critical role for system a amino acid transport in the regulation of dendritic development by BDNF. J Biol Chem 282: 5152 - 5159.
    Primary Dendritic Cells (Bone Marrow) Mouse GeneSilencer siRNA Transfection Reagent Chung, J-S, Sato, K., Dougherty, I.I., Cruz, Jr., P.D. and Ariizumi, K. (2007) DC-HIL is a negative regulator of T lymphocyte activation. Blood. 109(10): p. 4320-4327.
    Primary Cortical Neurons Mouse GeneSilencer siRNA Transfection Reagent Cui, H., Hayashi, A., Sun, H-S, Belmares, M.P., Cobey, C., Phan, T., Schweizer, J., Salter, M.W., Wang, Y.T., Tasker, R.A., Garman, D., Rabinowitz, J., Lu, P.S. and Tymianski, M. (2007) PDZ Protein Interactions Underlying NMDA Receptor-Mediated Excitotoxicity and Neuroprotection by PSD-95 Inhibitors. J. Neurosci. 27(37): 9901-9915.
    Primary Coronary Artery Endothelial Human HCAEC GeneSilencer siRNA Transfection Reagent Dandapat, A., Hu, C., Sun, L. and Mehta, J.L. (2007) Small Concentrations of oxLDL Induce Capillary Tube Formation From Endothelial Cells via LOX-1 Dependent Redox-Sensitive Pathway. Arterioscler Thromb Vasc Biol.
    Primary Peritoneal Macrophage Mouse GeneSilencer siRNA Transfection Reagent de Beer, M.C., Zhao, Z., Webb, N.R., van der Westhuyzen, D.R. and de Villiers, W.J.S. (2003) Lack of a direct role for macrosialin in oxidized LDL metabolism J. Lipid Res. 44: 674 - 685.
    Primary Dorsal Root Ganglion Neurons Mouse GeneSilencer siRNA Transfection Reagent Haruhisa Higuchi, Toshihide Yamashita, Hideki Yoshikawa, and Masaya Tohyama (2003) Functional inhibition of the p75 receptor using a small interfering RNA. Biochem. & Biophys. Res. Comm. 301: 804809.
    Primary Retinal Ganglion Rat GeneSilencer siRNA Transfection Reagent Hayashi, H., Campenot, R.B., Vance, D.E. and Vance, J.E. (2007) Apolipoprotein E-Containing Lipoproteins Protect Neurons from Apoptosis via a Signaling Pathway Involving Low-Density Lipoprotein Receptor-Related Protein-1. J. Neurosci. 27(8): 1933-1941.
    Primary Dorsal Root Ganglion Rat GeneSilencer siRNA Transfection Reagent Hengst, U. Cox, L.J., Macosko, E.Z. and Jaffrey, S.R. (2006) Functional and Selective RNA Interference in Developing Axons and Growth Cones. J. Neurosci. 26(21): 5727-5732.
    Primary Schwann Cells Mouse GeneSilencer siRNA Transfection Reagent Higuchi, H., Yamashita, T., Yoshikawa, H. and Tohyama, M. (2003) Functional inhibition of the p75 receptor using a small interfering RNA. Biochem. & Biophys. Res. Comm. 301: 804809.
    Primary Cardiac Fibroblast Mouse GeneSilencer siRNA Transfection Reagent Hu, C., Dandapat, A., Sun, L., Khan, J.A., Liu, Y., Hermonat, P.L. and Mehta, J.L. (2008) Regulation of TGFbeta 1-mediated collagen formation by LOX-1: Studies based on forced over-expression of TGFbeta 1 in wild-type and LOX-1 knockout mice cardiac fibroblasts. J. Biol. Chem. 283 (16) 10226-10231.
    Primary Cardiac Myocytes (Neonatal) Rat GeneSilencer siRNA Transfection Reagent Juhaszova, M., Zorov, D.B., Kim, S-H, Pepe, S., Fu, O., Fishbein, K.W., Ziman, B.D., Wang, S., Ytrehus, K., Antos, C.L., Olson, E.N. and Sollott, S.J. (2004) Glycogen synthase kinase-3{beta} mediates convergence of protection signaling to inhibit the mitochondrial permeability transition pore. J. Clin. Invest. 113(11): p. 1535-1549.
    Primary Lung Microvascular Endothelial Cells Human GeneSilencer siRNA Transfection Reagent Kolosova, I.A., Ma, S-F, Adyshev, D.M., Wang, P., Ohba, M., Natarajan, V., Garcia, J.G.N. and Verin, A.D. (2004) Role of CPI-17 in the regulation of endothelial cytoskeleton AJP: Lung. 287(5): L970.
    Primary Aortic Muscle Cells Rat GeneSilencer siRNA Transfection Reagent Lake, A.C. & Castellot, J.J. (2003) CCN5 modulates the antiproliferative effect of heparin and regulates cell motility in vascular smooth muscle cells. Cell Comm. & Signaling 1: 5.
    Primary Dendritic Cells Mouse GeneSilencer siRNA Transfection Reagent Li, M., Qian, H., Ichim, T.E., Ge, W-W, Popov, I.A., Rycerz, K., Neu, J., White, D., Zhong, R., Min, W-P. (2004) Induction of RNA Interference in Dendritic Cells. Immunologic Research 30 (2) 215-230.
    Primary Dendritic Cells Mouse GeneSilencer siRNA Transfection Reagent Li, M., Zhang, X., Zheng, X., Lian, D., Zhang, Z-X., Ge, W., Yang, J., Vladau, C., Suzuki, M., Chen, D., Zhong, R., Garcia, B., Jevnikar, A.M. and Min, W-P (2007) Immune Modulation and Tolerance Induction by RelB-Silenced Dendritic Cells through RNA Interference. J. Immunol. 178(9): p. 5480-5487.
    Primary Dendritic Cells Mouse GeneSilencer siRNA Transfection Reagent Liu, G., Ng, H., Akasaki, Y., Yuan, X., Ehtesham, M., Yin, D., Black , K.L. and Yu, J.S. (2004) Small interference RNA modulation of IL-10 in human monocyte-derived dendritic cells enhances the Th1 response. Eur. J. Immunol. 34: 1680-1687.
    Primary Cerebellar Neurons Rat GeneSilencer siRNA Transfection Reagent Numakawa, T., Nakayama, H., Suzuki, S., Kubo, T., Nara, F., Numakawa, Y., Yokomaku, D., Araki, T., Ishimoto, T., Ogura, A. and Taguchi, T. (2003) Nerve growth factor-induced glutamate release is via p75 receptor, ceramide and Ca2+ from ryanodine receptor in developing cerebellar neurons. J. Biol. Chem: 278: 41259-41269.
    Primary Osteoblast Mouse GeneSilencer siRNA Transfection Reagent Ohyama, Y., Nifuji, A., Maeda, Y., Amagasa, T. and Noda, M. (2004) Spaciotemporal Association and Bone Morphogenetic Protein Regulation of Sclerostin and Osterix Expression during Embryonic Osteogenesis. Endocrinology. 145(10): p. 4685-4692.
    Primary HMVEC Human GeneSilencer siRNA Transfection Reagent Radu Stefanescu, Dustin Bassett, Rozbeh Modarresi, Francisco Santiago, Mohamad Fakruddin, and Jeffrey Laurence (2008) Synergistic interactions between interferon-{gamma} and TRAIL modulate c-FLIP in endothelial cells, mediating their lineage-specific sensitivity to thrombotic thrombocytopenic purpura plasma-associated apoptosis. Blood 112: 340 - 349.
    Primary Cardiac Myocytes Feline GeneSilencer siRNA Transfection Reagent Ramabadran, R. S., Chancey, A., Vallejo, J.G., Barger, P.M., Sivasubramanian, N. and Mann, D.L. (2008) Targeted Gene Silencing of Tumor Necrosis Factor Attenuates the Negative Inotropic Effects of Lipopolysaccharide in Isolated Contracting Cardiac Myocytes. Tex Heart Inst J. 35(1): 16-21.
    Primary T-Cells (CD3+) Human GeneSilencer siRNA Transfection Reagent Samten, B., Howard, S.T., Weis, S.E., Wu, S., Shams, H., Townsend, J.C., Safi, H. and Barnes, P.F. (2005) Cyclic AMP Response Element-Binding Protein Positively Regulates Production of IFN-{gamma} by T Cells in Response to a Microbial Pathogen. J. Immunol. 174(10): p. 6357-6363.
    Primary Cortical Neurons Mouse GeneSilencer siRNA Transfection Reagent Sato, S., Xu, J., Okuyama, S., Martinez, LB., Walsh, S.M, Jacobsen, M.T, Swan, R.J., Schlautman, J.D., Ciborowski, P. and Ikezu, T. (2008) Spatial learning impairment, enhanced CDK5/p35 activity, and downregulation of NMDA receptor expression in transgenic mice expressing tau-tubulin kinase 1. J Neurosci 28(53): 14511.
    Primary Dendritic Cells Human GeneSilencer siRNA Transfection Reagent Smith, A.L., Ganesh, L., Leung, K., Jongstra-Bilen, J., Jongstra, J. and Nabel, G.J. (2007) Leukocyte-specific protein 1 interacts with DC-SIGN and mediates transport of HIV to the proteasome in dendritic cells. J. Exp. Med. 204(2): p. 421-430.
    Primary Cortical Neurons Mouse GeneSilencer siRNA Transfection Reagent Supnet, C., Grant, J., Kong, H., Westaway, D. and Mayne, M. (2006) Amyloid β-(1-42) Increases Ryanodine Receptor-3 Expression and Function in TgCRND8 Mice. J. Biol. Chem. 281: 38440 - 38447.
    Primary Aveolar Type II Cells Mouse GeneSilencer siRNA Transfection Reagent Ueno, T., Linder, S., Na, C-L, Rice, W.R., Johansson, J., and Weaver, T.E. (2004) Processing of Pulmonary Surfactant Protein B by Napsin and Cathepsin H. J. Biol. Chem. 279: 16178-16184.
    Primary Macrophage Mouse GeneSilencer siRNA Transfection Reagent Wang, Y., Chen, T., Han, C., He, D., Liu, H., An, H., Cai, Z. and Cao, X. (2007) Lysosome-associated small Rab GTPase Rab7b negatively regulates TLR4 signaling in macrophages by promoting lysosomal degradation of TLR4. Blood 110: 962 - 971.
    Primary Dorsal Root Ganglion Rat GeneSilencer siRNA Transfection Reagent Wu, K.Y., Hengst, U., Cox, L.J., Macosko, E.Z., Jeromin, A., Urquhart, E.R., and Jaffrey, S.R. (2005) Local translation of RhoA regulates growth cone collapse. Nature 436: 1020-1024.
    Primary Lacrimal Gland Acinar Cells Rabbit GeneSilencer siRNA Transfection Reagent Xie, J., Chiang, L., Contreras, J., Wu, K., Garner, J.A., Medina-Kauwe, L. and Hamm-Alvarez, S.F. (2006) Novel Fiber-Dependent Entry Mechanism for Adenovirus Serotype 5 in Lacrimal Acini. J. Virol. 80(23): 11833 - 11851.
    Primary Bone Marrow Mouse GeneSilencer siRNA Transfection Reagent Yang, R., Cai, Z., Zhang, Y., Yutzy IV, W.H., Roby, K.F. and Roden, R.B.S. (2006) CD80 in Immune Suppression by Mouse Ovarian Carcinoma驴Associated Gr-1+CD11b+ Myeloid Cells. Cancer Res 66 (13): 6807 - 6815.
    Primary Dendritic Cells Mouse GeneSilencer siRNA Transfection Reagent Zheng, X., Koropatnick, J., Li, M., Zhang, X., Ling, F., Ren, X., Hao, X., Sun, H., Vladau, C., Franek, J.A., Feng, B., Urquhart, B.L., Zhong, R., Freeman, D.J., Garcia, B. and Min, W-P. (2006) Reinstalling Antitumor Immunity by Inhibiting Tumor-Derived Immunosuppressive Molecule IDO through RNA Interference. J. Immunol. 177(8): 5639-5646.
    Primary Pulmonary Artery Endothelium Human GeneSilencer siRNA Transfection Reagent Zhuowei Li, Xhevahire Hyseni, Jacqueline D. Carter, Joleen M. Soukup, Lisa A. Dailey, and Yuh-Chin T. Huang (2006) Pollutant particles enhanced H2O2 production from NAD(P)H oxidase and mitochondria in human pulmonary artery endothelial cells. Am J Physiol Cell Physiol 291: C357 - C365.
    Primary HPAEC Pulmonary Artery Endothelium Human GeneSilencer siRNA Transfection Reagent Rentsendorj, O., Mirzapoiazova, T., Adyshev, D., Servinsky, L.E., Renne, T., Verin, A.D. and Pearse, D.B. (2008) Role of vasodilator-stimulated phosphoprotein in cGMP-mediated protection of human pulmonary artery endothelial barrier function. Am J Physiol Lung Cell Mol Physiol. 294(4): L686 -L697.
    RAW 264.7 Macrophage Mouse GeneSilencer siRNA Transfection Reagent de Beer, M.C., Zhao, Z., Webb, N.R., van der Westhuyzen, D.R. and de Villiers, W.J.S. (2003) Lack of a direct role for macrosialin in oxidized LDL metabolism. J. Lipid Res. 44: 674 - 685.
    RAW 264.7 Macrophage Mouse GeneSilencer siRNA Transfection Reagent Liu, X., Yao, M., Li, N., Wang, C., Zheng, Y. and Cao, X. (2008) CaMKII promotes TLR-triggered proinflammatory cytokine and type I interferon production by directly binding and activating TAK1 and IRF3 in macrophages. Blood. 112(13): 4961-4970.
    Renca Renal Cell Carcinoma Mouse GeneSilencer siRNA Transfection Reagent Ogushi, T., Takahashi, S., Takeuchi, T., Urano, T., Horie-Inoue, K., Kumagai, J., Kitamura, T., Ouchi, Y., Muramatsu, M. and Inoue, S. (2005) Estrogen Receptor-Binding Fragment-Associated Antigen 9 Is a Tumor-Promoting and Prognostic Factor for Renal Cell Carcinoma. Cancer Res. 65(9): p. 3700-3706.
    SaOs2 Osteosarcoma Human GeneSilencer siRNA Transfection Reagent Chu, F.,Chou, P.M., Zheng, X., Mirkin, B.L. and Rebbaa, A.驴(2005)驴Control of Multidrug Resistance Gene mdr1 and Cancer Resistance to Chemotherapy by the Longevity Gene sirt1. Cancer Res.65 (22): p. 10183-10187.
    SaOS2 Osteosarcoma Human GeneSilencer siRNA Transfection Reagent Zheng, X., Chu, F., Chou, P.M., Gallati, C., Dier, U., Mirkin, B.L., Mousa, S.A. and Rebbaa, A. (2009) Cathepsin L inhibition suppresses drug resistance in vitro and in vivo: a putative mechanism. Am J Physiol Cell Physiol. 296(1): C65-C74.
    Saos-2 Osteosarcoma Human GeneSilencer siRNA Transfection Reagent Zheng, X., Chou, P.M., Mirkin, B.L. and Rebbaa, A. (2004) Senescence-initiated Reversal of Drug Resistance: Specific Role of Cathepsin L. Cancer Res. 64: 1773 - 1780.
    SCCH196 Sarcoma Human GeneSilencer siRNA Transfection Reagent Takahashi, A., Higashino, F., Aoyagi, M., Yoshida, K., Itoh, M., Kyo, S., Ohno, T., Taira, T., Ariga, H., Nakajima, K., Hatta, M., Kobayashi, M., Sano, H., Kohgo, T. and Shindoh, M., (2003) EWS/ETS Fusions Activate Telomerase in Ewings Tumors. Cancer Res. 63: 8338-8344.
    SNB19 Glioblastoma Human GeneSilencer siRNA Transfection Reagent Song, S.W., Fuller, G.N., Zheng, H. and Zhang, W. (2005) Inactivation of the Invasion Inhibitory Gene IIp45 by Alternative Splicing in Gliomas. Cancer Res. 65(9): p. 3562-3567.
    SW480 Colon Adenocarcinoma Human GeneSilencer siRNA Transfection Reagent Zhu, S., Wang, W., Clarke, D.C. and Liu, X. (2007) Activation of Mps1 Promotes Transforming Growth Factor-β-independent Smad Signaling. J. Biol. Chem. 282(25): 18327-18338.
    U20S Osteosarcoma Human GeneSilencer siRNA Transfection Reagent Hotulainen P. and Lappalainen, P. (2006) Stress fibers are generated by two distinct actin assembly mechanisms in motile cells J. Cell Biol. 173(3): 383-394.
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    合肥基赛生物科技有限公司在发布的合肥基赛生物销售Crystalgen的0.1-10ul、1-200ul、100-1000ul、100-1300ul无DNA酶无RNA酶无热源、带滤芯吸头供应信息,浏览与合肥基赛生物销售Crystalgen的0.1-10ul、1-200ul、100-1000ul、100-1300ul无DNA酶无RNA酶无热源、带滤芯吸头相关的产品或在搜索更多与合肥基赛生物销售Crystalgen的0.1-10ul、1-200ul、100-1000ul、100-1300ul无DNA酶无RNA酶 查看更多>
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    问题补充:1.[单选题]与细菌侵袭力无关的酶是A.血浆凝固酶 B.氧化酶 C.DNA酶 D.卵磷脂酶E.透明质酸酶ABCDE 网友答案:参考答案:B 参考解析:氧化酶是细菌... 查看更多>
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    上海研生实业有限公司所提供的DNA酶甲基绿琼脂基础质量可靠、规格齐全,上海研生实业有限公司不仅具有精湛的技术水平,更有良好的售后服务和优质的解决方案,欢迎您来电咨询此产品具体参数及价格等详细信息! 查看更多>
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    第二章 DNA酶切及凝胶电泳第一节 概 述  一. DNA的限制性内切酶酶切分析   限制性内切酶能特异地结合于一段被称为限制性酶识别序列的DNA序列之内或其附近的特异位点上,并切割双链DNA。它可分为三类:Ⅰ类和Ⅲ类酶在同一蛋白质分子中兼有切割和 查看更多>
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    Taq DNA Polymerase 和rTaq酶有什么区别
    第一代热启动Taq酶是一种DNA聚合酶。 第一代指的是直接从Taq(这是一种细菌)体内提取的天然聚合酶,没有进行化学修饰的,为了达到更好的聚合催化效果,往往需要经过酶工程处理,得到的聚合酶催化效率更高。 热启动是指这种酶的反应温度条件
    Taq DNA polymerase是从克隆有Thermu aquaticus DNA Polymerase基因的大肠杆菌经诱导表达后,再经过柱纯化分离出来的一个约94 KD的重组蛋白。无外源核酸酶和细菌DNA污染,稳定性好,特异性强,适用于常规PCR扩增。
    DNA酶切及凝胶电泳 123
    挚爱小慧oDV2017-11-27
    限制性内切酶能特异地结合于一段被称为限制性酶识别序列的DNA序列之内或其附近的特异位点上,并切割双链DNA。它可分为三类:Ⅰ类和Ⅲ类酶在同一蛋白质分子中兼有切割和修饰(甲基化)作用且依赖于ATP的存在。Ⅰ类酶结合于识别位点并随机的切割识别位点不远处的DNA,而Ⅲ类酶在识别位点上切割DNA分子,然后从底物上解离。Ⅱ类由两种酶组成: 一种为限制性内切核酸酶(限制酶),它切割某一特异的核苷酸序列; 另一种为独立的甲基化酶,它修饰同一识别序列。Ⅱ类中的限制性内切酶在分子克隆中得到了广泛应用,它们是重组DNA的基础。向左转|向右转
    这种问题只能问佳学基因解码专家。
    基因克隆技术
    摘要:随着转基因技术的发展,获取目的基因片段的方法也越来越多样化,获取目的基因片段的技术也日新月异,但是无论获取的方法和手段怎么样发展,获取的主要思路始终在围绕一根主线在开展。
    关键词:基因 ,遗传效应,基因克隆,体外重组
    一、基因工程
    基因工程作为一门理论性与实践性较强的学科,其方法与技术已经渗透到现代生命科学的各个分支领域,成为生命科学的一门核心技术。基因工程包含许多独特的实验方法和技术,不仅内容丰富,涉及面广,实用性也强。基因工程是通过DNA 重组技术, 获得具有特殊生物遗传性状和功能的遗传工具生物体, 基因工程技术广泛应用于农业、医学、食品工业等。本文就基因工程的应用现状综合阐述。
    基因工程技术是一项极为复杂的高新生物技术, 它利用现代遗传学与分子生物学的理论和方法, 按照人类所需, 用DNA 重组技术对生物基因组的结构和组成进行人为修饰或改造, 从而改变生物的结构和功能, 使之有效表达出人类所需要的蛋白质或人类有益的生物性状。基因工程从诞生至今, 仅有30 年的历史, 然而, 无论是在基础理论研究领域, 还是在生产实际应用方面, 都已取得了惊人的成绩。首先,基因工程给生命科学自身的研究带来了深刻的变化。目前科学家已完成了多种细胞器的基因组全序列测定工作。其次, 基因工程具有广泛的应用价值, 能为工农业生产、医药卫生、环境保护开辟新途径。
    基因工程( 又称DNA 重组技术、基因重组技术) , 是20 世纪70 年代初兴起的技术科学, 是用人工的方法将目的基因与载体进行DNA重组, 将DNA 重组体送入受体细胞, 使它在受体细胞内复制、转录、翻译, 获得目的基因的表达产物。这种跨越天然物种屏障, 把来自任何生物的基因置于毫无亲缘关系的新的寄主生物细胞之中的能力, 是基因工程技术区别于其他技术的根本特征。 基因工程研究内容为:
    (1) 从复杂的生物有机体基因组中, 经过酶切消化或PCR 扩增等步骤, 分离出带有目的基因的DNA 片段。
    (2) 在体外, 将带有目的基因的外源DNA 片段连接到能够自我复制并具有选择记号的载体分子上, 形成重组DNA分子。
    (3)重组DNA 分子转移到适当的受体细胞, 并与之一起增殖。
    (4) 从大量的细胞繁殖群体中, 筛选出获得了重组DNA 分子的受体细胞克隆。
    基因工程中聚合酶不是基本酶的原因:DNA聚合酶是将单个的脱氧核苷酸加到原有的DNA片段上的,所以不用!DNA聚合酶是不论何种片段强制连接的,而聚合酶是连接有特定顺序的碱基对片段的DNA。基因工程常见几种酶比较:DNA聚合酶: 在DNA复制时起作用,将多个脱氧核苷酸催化聚合为脱氧核苷酸链(也就是DNA单链),此时形成的化学键是磷酸二酯键。DNA解旋酶: 在DNA复制或转录时起作用,将DNA的双链解开,作用对象是氢键,使氢键断裂 DNA水解酶: 在消化道或细胞内起作用,将DNA水解为脱氧核苷酸,作用对象是磷酸二酯键。DNA连接酶:在DNA修饰过程或基因工程中起作用,将两个DNA片段连接起来,此时形成的化学键是磷酸二酯键。
    解释酶的活性部位?必须基因及两者关系?
    目前常用方法有:化学修饰法、反应动力学法、X-射线衍射法等.1.化学修饰法:此方法是研究最早,应用最广泛的方法.原则上讲,酶分子侧链上的各种基团
    A、酶的本质是绝大多数酶是蛋白质,所以绝大多数酶是基因转录和翻译的产物,A错误; B、细胞核基因是染色体的组成部分,细胞质基因不是染色体的组成部分,酶不是细胞内染色体的组成成分,B错误; C、基因可以通过控制酶的合成控制代谢进而控制生物的性状,C正确; D、有某种酶的基因,细胞中不一定有相应的酶,因为在细胞中基因是选择表达的,D错误.故选:C.
    ‘控制’这个说法其实是个广泛的表述。实际上,DNA只不过是一个模板,通俗说是没有化学活性意义的母本,要转录翻译成为蛋白质,就要先转录成mRNA,然后mRNA作为最后的模板,翻译出想要得到的蛋白质,这之后,通常对得到的蛋白质进行进一步修饰才会得到最后由化学活性意义的蛋白质产物。当然这个步骤很复杂,远远不是说得这么简单,讨论他们都是化学分子组成的同类这一点没有意义的,就像你的手的行为有大脑控制的,而你的手和大脑都是蛋白质组成的。。。这个逻辑关系实际是宏观和微观的关系,朋友千万不要把这些混淆了去钻牛角尖。如果有兴趣和能力学量子物理,你就都明白了。。。祝学习愉快。
    核酸酶发展历史123
    TSˊ控白2021-07-21
    20世纪70年代,在细菌中陆续发现了一类核酸内切酶,能专一性地识别并水解双链DNA上的特异核苷酸顺序,称为限制性核酸内切酶(restriction endonuclease,简称限制酶)。当外源DNA侵入细菌后,限制性内切酶可将其水解切成片段,从而限制了外源DNA在细菌细胞内的表达,而细菌本身的DNA由于在该特异核苷酸顺序处被甲基化酶修饰,不被水解,从而得到保护。
    限制性核酸内切酶的研究和应用发展很快,已提纯的限制性核酸内切酶有100多种,许多已成为基因工程研究中必不可少的工具酶。
    限制性核酸内切酶可被分成三种类型。Ⅰ型和Ⅲ型限制酶水解DNA需要消耗ATP,全酶中的部分亚基有通过在特殊碱基上补加甲基基团对DNA进行化学修饰的活性。
    Ⅰ型和Ⅲ型酶具有限制和修饰两种作用,而特异性弱,切割位点的序列不固定,不已知,不宜用于基因克隆中。
    Ⅱ型限制酶水解DNA不需要ATP也不以甲基化或其它方式修饰DNA,能在所识别的特殊核苷酸顺序内或附近切割DNA。因此,被广泛用于DNA分子克隆和序列测定。
    然后如何去除DNA中的RNA酶 123
    禽兽TA02072017-11-27
    (一)从源头控制污染:
    (1)玻璃制品、塑料制品和电泳槽 灭菌的一次性使用的塑料制品基本上无RNA酶,可以不经预处理直接用于制备和贮存RNA。实验室用的普通玻璃器皿和塑料制品经常有RNA酶法染,使用前必须于180 ℃干烤8小时或更长时间(玻璃器皿)或用氯仿冲洗(塑料制品)。另一种方法是用0.1 %焦碳酸二乙酯(DEPC)的水溶液浸泡用于制备RNA的烧杯,试管和其他用品。DEPC是RNA酶的强烈抑制剂,但其作用并不是绝对的(Fedorcsak和Ehrenberg,1966)。灌满DEPC的玻璃或塑料器皿在37℃放置2小时,然后用灭菌水淋洗数次, 并于100℃干烤15分钟(Kumar和Lindberg,1972)。在15 lbf/in2(1.034x105Pa)高压蒸氯灭菌15分钟。上述处理可以除去器甲上痕量的DEPC,以防DEPC通过羧甲基化作用对RNA的嘌呤碱基进行修饰。
    用于RNA电泳的电泳槽应用去污剂洗干净,再用水冲洗,用乙醇干燥,然后灌满3%的H2O2溶液,于室温放置10分钟,然后用0.1 %DEPC处理过的水彻底冲洗电泳槽。最好能留出一些玻璃器皿、塑料制品和电泳槽作上特殊标记,存放在指定地点,为RNA实验专用。
    (2)研究人员造成的污染 RNA酶最主要的潜在污染源是研究人员的手。因此,在准备分离的和分析RNA的材料和溶液时,主有涉及RNA的一切操作过程中,都应戴一次性手套,接触“胖的”玻璃器皿和其他物品以后,手套就可能沾染上RNA酶,因此进行RNA实验时应勤换手套。
    (3)污染的溶液 用高压灭菌的水和RNA研究专用的化学试剂配制溶液,用干烤过的药匙称取试剂,将溶液装入无RNA酶的玻璃器皿。可能的话溶液均应用0.1%DEPC于37℃至少处理12小时,然后于100℃加热15分钟或在15lbf/in2(1.034x105Pa)的高压下蒸气灭菌15分钟。注:DEPC可与胺类迅速发生化学反应,因些不能用来处理含有Tris 一类的缓冲液。可存几瓶新的未开封的Tris晶体以制备无RNA酶的溶液。
    (二)已经污染,可以加入RNA酶的抑制剂
    (1)RNA酶的蛋白质抑制剂是 从人胎盘分离的一种蛋白质可与多种RNA酶紧密结合(KI≈3x1010)形成非共价结合的等摩尔复合物,使RNA酶失活。此蛋白质体内可能是血管生成素的抑制剂,血管生成素是氨基酸序列和推测的三级结构与胰RNA酶类似的一种血管生成因子, 几个厂家以不同的商品名出售这种抑制剂,该蛋白质应置于含5mmol/L二硫苏糖醇(DTT)的50%甘油中,贮存于-20℃。抑制剂制品冻融数次后或放置在氧化条件下即应弃之不用,因为上述处理会使蛋白质变性从而释放出所结合的RNA酶。因此,在使用变性剂裂解哺乳动物(mammal;mammalian)细胞这一提取RNA的初始步聚中不应使用这种蛋白抑制剂。然而职用更温和的裂解方法时应使用这种抑制剂,并且在后续的所有RNA纯化步骤中均应有此蛋白质存在。由于酚抽提可以除蛋白质抑制剂,故应在纯化过程中补加几次抑制剂。其最大活性的发挥要求巯基试剂,而且它并不干扰反转录或mRNA在无细胞体系中的翻译。
    (2)氧钒核糖核苷复合物 这种由氧钒(1V)离子和4种核糖核苷之中的任意一种所形成的复合物,是量种过渡态类似物,它能与多种RNA酶结合并几科能百分之百地抑制RNA酶的活性。这4种氧钒核糖核苷复合物可加入完整细胞中,在RNA提取和纯化的所有过程中,其使用浓度都是10mmol/L。所得到的mRNA可直接在硅卵母细胞中进行翻译, 并能作为某些外酶促反应(如mRNA反转录)的模板。然而氧钒核糖核苷复合物强烈抑制mRNA在无细胞体系中的翻译,因此必须用含0.1 %羟基喹淋的苯酚[ 用0. 01mol/LTris.Cl(pH7.8)平衡]多次抽提以去除之。有几家公司出售氧钒核糖核苷复合物。
    (3)Macaloid(硅藻上)Macaloid是一咱粘土,很多年前就发现它能吸附RNA酶,用缓冲液将其制成浆液,以0.015%(W/V)的终浓度溶解细胞。 这种粘土随同它所吸附的RNA酶可在后续的RNA纯化过程中(如酚抽提后)经离心去除。
    (三)提取核酸时可以加入盐酸胍或硫氰酸胍溶液。盐酸胍或硫氰酸胍溶液能迅速溶解蛋白质,从而免受RNA酶的干扰。
    作为生物体四大有机分子之一,糖的代谢是整个生物代谢的中心。它既是植物的生长发育过程中重要的结构组分,又是一种细胞间用于通讯的信号分子。糖类的代谢主要包括合成、修饰、分解等过程,参与这些过程的酶根据功能不同各司其职。如糖类的合成需要糖基转移酶(glycosyl transferases);糖类的修饰需要糖脂酶(carbohydrate esterases);分解则需要糖苷水解酶和多糖裂解酶(glycoside hydrolases and polysaccharide lyases)等。 糖苷水解酶(glycoside hydrolase,GH)亦称为糖苷酶,是作用于各种糖苷或寡糖使糖苷键水解的酶类总称。根据氨基酸序列,可以将糖苷水解酶划分为130个家族(GHs),但是至今仍有很多基因没有被分到任何一个家族中。拟南芥基因组中已鉴定出401个糖苷水解酶基因,分别属于35个不同家族。糖苷水解酶作为糖类代谢的关键组分,其生物学功能得到了广泛的研究。 本文利用反向遗传学,借助遗传转化和RT-PCR等生化和分子生物学手段,研究了拟南芥中两个新的类糖苷酶基因AtGHL1和AtGHL2的生物学功能。研究发现: 1.通过生物信息学分析,AtGHL1和AtGHL2两个基因的核苷酸及氨基酸序列同源性达90%以上,其编码蛋白均有糖苷酶特有的DUF1680复合结构域和类6-发卡糖苷酶,但这两个基因没有被归属到目前分类的任何糖苷酶家族中,为两个新型的类糖苷水解酶。 2.通过PCR鉴定及RT-PCR检测,分别获得了AtGHL1和AtGHL2的基因缺失突变体ghl1和ghl2;通过RNAi载体的构建及遗传转化,获得两个基因表达均下调的转基因植株ghli。表型检测发现,各突变体均存在雄蕊缺失的现象,表明AtGHL基因可能参与了雄蕊发育的过程。 3.根据拟南芥遗传转化株中报告基因GUS和YFP的表达,发现这两个基因主要在幼苗、莲座叶及花等器官的输导组织中表达。亚细胞定位结果显示,AtGHL定位于细胞壁。 4.通过糖饥饿胁迫及RT-PCR检测,发现AtGHL基因转录受糖饥饿的诱导。在不含蔗糖的MS培养基中,突变体ghl1和ghli幼苗生长受到严重影响,存活率仅为20%。
    限制性核酸内切酶(以下简称限制性酶)是一类识别双链DNA中特定核苷酸序列的DNA水解酶,以内切方式水解DNA,产生5’-P和3’-OH末端。  1952年Luria等及1953年Bertani等研究噬菌体时发现了宿主控制性现象。Arber及其同事用放射性同位素标记证明,噬菌体在新品系中的损害伴随有其DNA的降解,但宿主自己的DNA并不降解,据此他们提出了限制 - 修饰酶假说。对于一个宿主细胞,限制性酶及 DNA甲基化酶是其细胞中的一对酶,它们对DNA底物有相同的识别顺序,但有相反的生物功能,限制性酶的功能是在DNA分子内部拆卸水解,甲基化酶是修饰,DNA分子经修饰后,就可逃避限制性酶的识别,而甲基化酶只修饰宿主自身的DNA,从而避免了限制性酶对自身DNA的破坏。
    限制性酶主要分为三种类型:Ⅰ型限制酶为复合功能酶,具有限制-修饰两种功能,但在 DNA链上没有固定的切割位点,一般在离切割位点1kb到几kb的地方随机切割,不产生特异性片段。Ⅲ型酶与Ⅰ型酶基本相似,不同的是Ⅲ型酶有特异性的切割位点,但这两类酶对 DNA酶切分析的意义不大,通常所说的限制性内切酶是指Ⅱ型酶,它能够识别与切割DNA链上的特定的核苷酸顺序,产生特异性的DNA片段。
    2.识别序列及消化产物的末端结构 限制性酶的识别序列,大部分具有双轴对称性结构或称回文序列,如EcoRI的识别序列为:
    GAA
    TTC
    横轴
    CTT
    AAG
    纵轴
    将纵轴一侧的序列以横轴为中心旋转180°,则纵轴两侧的序列相互对称,这种结构又称为双重对称结构。大部分酶的识别序列长度为4-6个核苷酸。4核苷酸序列在DNA链中出现频率高,对一随机排列的DNA分子来说,理论值为1/44,因此4核苷酸识别序列的限制性酶在DNA链上切点多,产生片段的数目多,长度短,显示出酶的特异性较低。对于5和6核苷酸识别序列的酶,出现频率分别为1/45和1/46 ,因此,6核苷酸序列在DNA中出现频率低,酶的特异性强,而8核苷酸识别位点在DNA链中出现机率更低(1/48 ),特导性更强,可提供更长的DNA片段。一部分限制性酶具有非典型的双轴对称性序列,其回文识别序列被个或几个其他核苷酸所间隔,如BglⅠ,这种酶的特异性比识别长度相同的典型回文序列的酶略高。另外有些限制性酶(约10种,如BbVⅠ等),其识别序列不表现为回文结构,它们降解双链DNA时,酶切点大部分不在识别序列内,而是与识别序列相距5至13个核苷酸残基不等。
    限制性酶切片段的末端结构:限制性酶不但有特定的识别序列,并且任何一种酶切割 DNA链时,总是水解核苷酸3’和5’-磷酸二酯键的3’位磷酸酯键,使产物的5’端带磷酸单酯基团,而3’末端则为游离羟基。因此某一种酶的全部产物的末端具有相同的结构。根据切点序列的结构特点,产物的末端可分为粘性末端和平末端两类。粘性末端指酶切后DNA片段末端带有1-4个核苷酸残基的单链结构,而片段两端突出的单链具有互补性,突出的单链因部位的不同,又可分为5’-与3’-粘性末端两种,突出的单链带5’磷酸单酯的称5’-粘性末端,而突出的单链含3’-羟基则称3’-粘性末端。平末端指酶切后,片段为齐头末端结构。在DNA体外重组时,粘性末端是DNA连接酶的有效底物,有很高的连接效率。向左转|向右转