说明书(![](\"http://cache1.bioon.com.cn/ewebeditor/sysimage/icon16/pdf.gif\")
bmeg_handbook.pdf)Features:Stable, high-yield protein expression in Bacillus megaterium Ideal for both small and large-scale protein production Tightly regulated and efficiently inducible xylose operon (up to 350-fold) No alkaline proteases activity even up to 5 hours after induction No endotoxins observed Versatile cloning (extended polylinker, additional BsrG I site*) Versatile production (intracellular or extracellular) Versatile purification (native, 6xHis-tag, Strep-tag, Strep/6xHis double-tag) Removeable tag versions (for TEV or Xa proteases) available ![\"New!\"](\"http://www.mobitec.com/bilder/new.gif\")
Compatible with all B. subtilis vectors
*BsrG I site is not included in all vectors.
The B. megaterium expression system provides a versatile and easy-to-handle tool for stable and high-yield protein production, both small and large scale.
B. megaterium has proven to be an excellent alternative host to E. coli for heterologous gene expression. Unlike other bacilli strains, proteolytic degradation by alkaline proteases is avoided. In addition, there are no endotoxins found in the cell wall.
High Protein YieldAs a result, protein yields are exceptionally high even if inexpensive substrates are used. For example, the proteins mutarotase (Mro) and glucose dehydrogenase (Gdh) have been accumulated up to 20% and 30% of the total soluble protein, respectively. Using the tightly regulated xylose operon, the genes were induced 130- to 350-fold without proteolysis.
Versatile System with a Wide Range of VectorsMoBiTec provides a wide range of useful vectors for the B. megaterium system that are adaptable for most applications and protein purification methods. These include vectors carrying a secretion signal peptide sequence, a 6xHis-tag, a Strep-tag, and a Strep/6xHis double-tag. Additionally, all vectors contain the tightly regulated and highly inducible xylose operon promoter.
B. megaterium protoplasts specifically optimized for transformation The protoplasts supplied by MoBiTec are from B. megaterium strain WH320 developed by Prof. Dr. W. Hillen at the Institute of Microbiology in Erlangen, Germany. These protoplasts are prepared according to an optimized protocol resulting in the highest transformation efficiencies. For secretion vectors we offer protoplasts of strain MS941. Both strains are mutants of DSM319, where WH320 is a chemically mutant, while MS941 has a defined deletion in the gene of the major extracellular protease NprM.
Examples:Selected Proteins successfully over-produced in Bacilli strains with our B. megaterium vectors:
β-Galactosidase (LacZ)1 Catabolite control protein (CcpA)2,3 Clostridium difficile toxin A4 Cobaltochelatase (CbiX)5 Dextransucrase6 Endolevanase (LevB)7 Glucose dehydrogenase (GdhA)1 Heat shock protein (HPr) from PTS (phosphotransferase sugar transport system)8 Human single-chain urokinase-like plasminogen activator (rscuPA)1 Levansucrase9 Mutarotase (Mro)1 Neopullulanase10 Translocation ATPase of the preprotein translocase (SecA)11 Trehalose repressor (TreR)12 Bacillus megaterium protoplasts ready for transformation (strain WH320;1)Material is sufficient for 4 transformations plus control experiment.5 x 500 lBMEG31pC-His1623hp shuttle vector, C-term. 6xHis-tag; lyophilized DNA10 μgBMEG32pN-His-TEV1623hp shuttle vector, N-term. His-tag,TEV-site10 μgBMEG33pSPLipA-hp, shuttle vector with signal sequence, lypophilized DNA10 μgBMEG34pSPYocH-hp, shuttle vector with signal sequence, lypophilized DNABMEG03pWH1520 shuttle vector, original; lyophilized DNABMEG10pMM1522 shuttle vector, improved; lyophilized DNABMEG11pMM1525 shuttle vector with signal sequence; lyophilized DNABMEG12pHIS1522 shuttle vector, 6xHis-tagged; lyophilized DNA precursor of BMEG20 10 g BMEG13pHIS1525 shuttle vector with signal sequence; 6xHis-tagged; lyophilized DNABMEG14pSTREP1525 shuttle vector with signal sequence; Strep-tagged; lyophilized DNABMEG15pSTREPHIS1525 shuttle vector with signal sequence; Strep/6xHis double-tagged; lyophilized DNA BMEG20pC-His1622 shuttle vector, C-term. 6xHis-tag; lyophilized DNA 10 g BMEG21pC-Strep1622 shuttle vector, C-term. Strep-tag; lyophilized DNA BMEG22pN-His-TEV1622 shuttle vector, N-term. His-tag,TEV-site BMEG23pN-Strep-TEV1622 shuttle vector, N-term. Strep-tag,TEV-site BMEG24pN-StrepXa1622 shuttle vector, N-term. Strep-tag; Xa site; lyophilized DNA BMEGT702Bacillus megaterium protoplasts, strain MS941, pretransformed with pT7-RNAP5 x 500 LBMEGT701Bacillus megaterium high yield T7 gene expression kit, includes pretransformed protoplasts BMEGT702 (5 x 500 l), pPT7 cloning vector and pPT7-GFP control vector (vectors lyophilized, 10 g each) BMEGT710Bacillus megaterium pPT7 cloning vector, lyophilizedshipped at RT, protoplasts shipped on dry ice store lyophilized vectors at 4°C, reconstituted vectors at -20°C, protoplasts at -70°CAll vectors are E. coli / B. megaterium shuttle vectorsPlease note that protoplasts of Bacillus megaterium have a limited shelf life of some months only, depending on the date of production. Therefore purchase and use of protoplasts must be thoroughly planned.蚂蚁淘在线声明：以上所展示的信息由企业自行提供，内容的真实性、准确性和合法性由发布企业负责。蚂蚁淘在线对此不承担任何保证责任。