Optimization of eGFP expression using a modified baculovirus expression system.
The baculovirus gene expression system is an efficient and safe protein expression system, since baculoviruses cannot replicate in mammalian cells. In order to improve the transduction efficiency and increase the reporter gene expression levels delivered by baculoviruses, we tested in the baculovirus expression cassette the Woodchuck hepatitis virus response element (WPRE), and AAV-derived inverted terminal repeats (ITRs) and the truncated vesicular stomatitis virus G protein (VSV-GED). The results showed that WPRE and VSV-GED have synergistic effects and could enhance the expression efficiency of enhanced green fluorescence protein (eGFP), and that ITRs effectively extended the duration of eGFP expression. We also demonstrated that the efficiency of eGFP expression varied under the control of the CMV, CBA, EF1-α or WSSV ie1 promoters in different cell lines.
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China. Electronic address: gejingping@126.com.
Jingping
J
Ge
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China.
Liying
L
Jin
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China.
Xiaoyan
X
Tang
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China.
Dongni
D
Gao
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China.
Qi
Q
An
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China.
Wenxiang
W
Ping
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D016466:CHO Cells; D002460:Cell Line; D002642:Chick Embryo; D003412:Cricetulus; D004279:DNA, Viral / Q000235:genetics*; D000229:Dependovirus / Q000235:genetics; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D020218:Response Elements; D061987:Sf9 Cells; D018411:Spodoptera; D020079:Terminal Repeat Sequences; D014161:Transduction, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics
D004279:DNA, Viral; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
ITRs; Recombinant baculovirus; VSV-GED; WPRE; WSSV ie1 promoter