A TRANSIENT 3-PLASMID EXPRESSION SYSTEM FOR THE PRODUCTION OF HIGH-TITER RETROVIRAL VECTORS

被引：612

作者：

SONEOKA, Y ^{[1
]}

CANNON, PM ^{[1
]}

RAMSDALE, EE ^{[1
]}

GRIFFITHS, JC ^{[1
]}

ROMANO, G ^{[1
]}

KINGSMAN, SM ^{[1
]}

KINGSMAN, AJ ^{[1
]}

机构：

[1] UNIV OXFORD,DEPT BIOCHEM,RETROVIRUS MOLEC BIOL GRP,OXFORD OX1 3QU,ENGLAND

来源：

NUCLEIC ACIDS RESEARCH | 1995年 / 23卷 / 04期

关键词：

D O I：

10.1093/nar/23.4.628

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have constructed a series of MLV-based retroviral vectors and packaging components expressed from the CMV promoter and carried on plasmids containing SV40 origins of replication, These two features greatly enhanced retroviral gene expression when introduced into cell lines carrying the SV40 large T antigen, The two packaging components, gag-pol and env, were placed on separate plasmids to reduce helper virus formation, Using a highly transfectable human cell line and sodium butyrate to further increase expression of each component, we achieved helper-free viral stocks of similar to 10(7) infectious units/ml by 48 h after transient co-transfection with the three plasmid components, This system can be used both for the generation of high titer retroviral stocks for transduction and for the rapid screening of a large number of MLV gag-pol or env mutants.

引用

页码：628 / 633

页数：6

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