Efficient gene delivery into mammalian cells by recombinant baculovirus containing a hybrid cytomegalovirus promoter/Semliki Forest virus replicon.
BACKGROUND
Baculovirus, which is widely utilized as an excellent tool for the production of recombinant protein in insect cells, has recently emerged as a novel and attractive gene delivery vehicle for mammalian cells. Alphavirus, such as Semliki Forest virus (SFV), has also received considerable attention for use as expression vectors because of its self-replicating property. In the present study, we investigated the characterization of recombinant baculovirus incorporating a hybrid cytomegalovirus (CMV) promoter/SFV replicon.


METHODS
Recombinant baculovirus containing the hybrid CMV promoter/SFV replicon was constructed. Using enhanced green fluorescence protein (EGFP) as the reporter gene, gene delivery efficiencies and the ability to express heterogenous protein in mammalian cells were evaluated. Optimal transduction conditions, including transduction temperature, time and dose, were also investigated.


RESULTS
The obtained recombinant baculovirus, Bac-CMV/SFV-EGFP, exhibited high transduction efficiency and high-level expression of reporter protein in mammalian cells. Furthermore, this recombinant baculovirus could induce apoptosis in mammalian cells in the course of transduction, as demonstrated by the observed DNA laddering patterns and increased caspase-3 activity.


CONCLUSIONS
The developed baculovirus vector has a high transduction efficiency and the ability to mediate foreign gene expression in mammalian cells. Taken together with its pro-apoptotic properties, this baculovirus vector may provide an alternative tool for vaccine development.
Pan
Yongfei
Y
Division of Animal Infectious Diseases, State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.
Zhao
Qian
Q
Fang
Liurong
L
Luo
Rui
R
Chen
Huanchun
H
Xiao
Shaobo
S
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003587:Cytomegalovirus / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D011401:Promoter Regions, Genetic; D012093:Replicon / Q000235:genetics*; D012672:Semliki forest virus / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D014162:Transfection
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins