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终止子 | SV40 poly(A) signal |
质粒大小 | 34947bp |
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
9.2-100是一个腺病毒骨架质粒,用于产生腺病毒,pacAd5 9.2-100质粒的参考资料是:RAPAd Universal Adenoviral Expression
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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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