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您现在的位置:
启动子 | CMV |
复制子 | pBR322 ori |
终止子 | SV40 poly(A) signal |
质粒大小 | 6215bp |
原核抗性 | Amp |
克隆菌株 | Stbl3 |
培养条件 | 37℃ |
质粒宿主 | 哺乳细胞,腺病毒 |
质粒用途 | 蛋白表达 |
原核抗性 | 氨苄青霉素 |
pacAd5 CMVK-NpA质粒是一个腺病毒质粒,CMV启动子驱动目的基因的表达。具体可点击查看该资料:RAPAd
CMV Adenoviral Expression System。
Recombinant adenoviruses have tremendous potential in both
research and therapeutic applications. There are numerous advantages they
provide when introducing genetic material into host cells. The permissive
host cell range is very wide. The virus has been used to infect many mammalian
cell types (both replicative and non-replicative) for high expression of the
recombinant protein. Recombinant adenoviruses are especially useful for
gene transfer and protein expression in cell lines that have
low transfection efficiency with liposome. After entering cells, the virus
remains epichromosomal (i.e. does not integrate into the host chromosome
so does not activate or inactivate host genes). Recently, recombinant
adenoviruses have been used to deliver RNAi into cells.
Two methods have traditionally been used to generate
recombinant adenoviruses. The first involves homologous recombination of a
shuttle vector containing gene of interest and an adenoviral
backbone plasmid vector (restricted in E1/E3) in an adenovirus packaging
cell line. The isolation of recombinant adenovirus by this method involves
performing multiple plaque isolations to avoid wild-type virus and is
extremely laborious and time consuming. The second method, pAdEasy system,
employs the homologous recombination machinery in E. coli, a recombinant
adenovirus is produced by a doublerecombination event between
cotransformed adenoviral backbone plasmid vector and a shuttle
vector carrying the gene of interest. For the pAdEasy method, the system
is high fidelity, but inefficient and requires the screening of many
bacterial colonies. This results in a significant time commitment
even before transfection of recombinant DNA into E1-expressing cells such
as HEK293 cells.
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