[Construction of BV-T7 hybrid expression system based on baculovirus to express target gene eGFP in mammalian and chicken cells].
OBJECTIVE
To investigate whether the recombinant baculovirus BV-T7 hybrid expression system can be effectively transduced into chicken cells in vitro, and whether it can express the foreign genes (eGFP).


METHODS
We established a hybrid baculovirus-T7 RNA polymerase system for transient expression in mammalian cells and chicken cells. Two recombinant baculoviruses were constructed, one carrying cDNA of bacteriophage T7 RNA polymerase, with a nuclear localization signal, under the control of a mammalian promoter and the other expressing eGFP gene under the control of T7 promoter. The constructed recombinant baculoviruses co-infected mammalian oligodendrocyte cells, as well as chicken embryo fibroblasts cells and chicken embryo skeletal muscle cells.


RESULTS
The eGFP activity was detected in mammalian cell lines and embryo fibroblasts cells and chicken embryo skeletal muscle cells. The recombinant baculovirus transduction efficiency of oligodendrocyte cells was 59.5%, and in CEF cells and myoblast cells the transduction efficiencies were 23.2% and 33.1%.


CONCLUSIONS
BV-T7 hybrid expression could be expressed T7 RNAP in mammalian cells and avian cells.
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, China. gejingping@126.com
Jingping
J
Ge
Xiaoyan
X
Tang
Dongni
D
Gao
Shanshan
S
Song
Shan
S
Lu
Zhuangwei
Z
Lou
Wenxiang
W
Ping
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002642:Chick Embryo; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D018390:Gene Targeting / Q000295:instrumentation* / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008322:Mammals / Q000235:genetics; D018411:Spodoptera; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases