Baculovirus-mediated gene expression in chicken primary cells.
A recombinant baculovirus was constructed containing an expression cassette with a reporter gene, green fluorescent protein, directed by a constitutive mammalian promoter: a human cytomegalovirus immediate early promoter/enhancer (CMV-IE). High titer virus was prepared with ultracentrifugation. Efficient gene delivery and expression were observed in the virus-treated chicken primary culture, myoblast cells, and whole embryonic fibroblast cells. It was noticed that an addition of sodium butyrate (a selective histone deacetylase inhibitor) to viral transduction medium extremely enhanced the reporter-gene expression. However, there is no effect of presence of trichostatin A observed. To maximize the reporter-gene expression, the baculoviral infection condition was optimized with both cell types. Our approaches demonstrated that recombinant baculovirus could efficiently deliver its genome DNA into chicken primary cells and that CMV-IE, a mammalian-cell-active promoter, was functional in chicken primary cells and could direct a high level of gene expression. Clearly, the recombinant baculovirus provides an alternative means for foreign gene delivery into avian cells.
Ping
Wenxiang
W
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, 74 Xuefu Road, Harbin, Peoples' Republic of China.
Ge
Jingping
J
Li
Shengxian
S
Zhou
Hui
H
Wang
Kun
K
Feng
Yonggang
Y
Lou
Zhuangwei
Z
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000502:physiology; D002478:Cells, Cultured; D002645:Chickens / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D049452:Green Fluorescent Proteins; D032446:Myoblasts / Q000378:metabolism*; D014170:Transformation, Genetic
D049452:Green Fluorescent Proteins