[Expression of target gene in eukaryotic cells driven by prokaryotic T7 promoter and its RNA polymerase].
To enhance the efficiency of the expression of target gene in eukaryotic cells, one of the strongest prokaryotic expression systems, the T7 RNA polymerase and T7 promoter, was introduced into eukaryotic cells. A duel-plasmid gene expression system of T7 bacteriophage components was developed; one containing the T7 phage RNA polymerase gene under the control of eukaryotic promoter CMV (pCMV-T7pol) and the other (pT7IRES) containing the T7 promoter and T7 terminator as well as EMCV IRES. To test the feasibility of this plasmid system for eukaryotic expression, hepatitis B virus envelop HBV preS2/S was used to construct pT7IRES-HBs. The target genes were expressed efficiently by the eukaryonized prokaryotic expression system in a variety of the cells indicating C2C12, SP2/0, NIH3T3 and BALB/c 3T3, suggesting the potential applications of the expression system in gene therapy and gene immunization.
Yuan
Zhi-Gang
ZG
Department of Immunology of Shanghai Medical College of Fudan University, Key Laboratory of Molecular Medicine of Ministry of Education, Center for Gene Immunization and Vaccine Research, Shanghai 200032, China.
Zhang
Jin-Ping
JP
Chu
Yi-Wei
YW
Wang
Ying
Y
Xu
Wei
W
Xiong
Si-Dong
SD
D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics*; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D018390:Gene Targeting; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases