주문정보
- - 재고수량은 변동될 수 있습니다.
- - 재고 확인 시 "갱신" 버튼을 누르시면 실시간 재고를 확인하실 수 있습니다.
- - 가격이 ‘별도문의‘ 시, 상단 ‘견적신청’ 버튼을 눌러 문의해주시면 빠른 답변을 받으실 수 있습니다.
선택 | Cat.No. | 제품명 | 가격(VAT별도) | 수량 |
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제품특징
□ pAcGFP1 Vector Information
□ Restriction map and multiple cloning site (MCS) of pAcGFP1.
Unique restriction sites are in bold. NOTE: The Xba I and Bcl I sites are methylated in the DNA provided by Clontech Laboratories, Inc. If you wish to digest the vector with these enzymes, you will need to transform the vector into a dam -host and make fresh DNA.
□ Location of features
● lac promoter: 95 - 178 CAP binding site: 111 - 124 - 35 region: 143 - 148;- 10 region: 167 - 172 Transcription start point: 179 lac operator: 179 - 199
● lacZ - ECFP fusion protein expressed in E. coli Ribosome binding site: 206 - 209 Start codon (ATG): 217 - 219; stop codon: 1006 - 1008
● 5" multiple cloning site: 234 - 281
● Aequorea coerulescens Green fluorescent protein (AcGFP1) gene Kozak consensus translation initiation site: 282 - 292 Start codon (ATG): 289 - 291; stop codon: 1006 - 1008 Insertion of Val at position 2: 292 - 294 Last amino acid: 1003 - 1005
●3" Multiple Cloning Site: 1009 - 1108Ampicillin resistance gene: Promoter:- 35 region: 1484 - 1450,- 10 region: 1507 - 1512 Ribosome binding site: 1542 - 1546 β-lactamase coding sequences:start codon (ATG): 1554 - 1556; stop codon: 2412 - 2414 β-lactamase signal peptide: 1554 - 1622 β-lactamase mature protein: 1623 - 2411
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由于质粒的不兼容性,拥有同种复制子的质粒不能在同一细胞内稳定共存,经过几代的复制,会质粒丢失,所以并不是任何两个或两个以上质粒都可以在同一细胞内稳定存在,但是可以同时进入.
稳定转染的细胞株,就是转染后质粒可以稳定整合到基因组上不会因细胞分裂而丢失。区别于质粒瞬时转染不能长时间保留质粒在细胞里。
如果经费充足的话可以找公司包装病毒,如武汉的普健可以提供各种载体的构建及细胞株构建的技术服务。
先提取RNA,反转录成cDNA,然后根据目的基因设计PCR引物,通过半定量RT-PCR确定目的基因表达.
正因为检测的是mRNA,所以要先反转录成cDNA才能PCR.
2.将构建完成的载体与慢病毒包装质粒混合,共转染靶细胞
3.收集病毒液
4.用病毒液感染靶细胞
5.用载体上带的抗生素进行筛选,如果没有,可以用无限稀释法
6.获得稳转株
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