title: Efficient gene delivery into fish cells by an improved recombinant baculovirus.
abstract: Studies on transduction of mammalian cells have shown that baculovirus can be used as an effective vector for gene delivery. However, previous studies have found the gene delivery efficiency to be very low in differentiated fish cells. In this study, an improved recombinant baculovirus, containing cytomegalovirus immediate-early (CMV-IE) promoter and an enhanced green fluorescent protein (EGFP) gene as the reporter gene, was constructed. The transduction efficiency of recombinant baculovirus in several fish cell lines was measured by flow cytometry (FCM) and the persistence of EGFP was monitored by fluorescence microscopy. The results demonstrated that baculovirus can mediate the high efficiency of gene delivery into differentiated fish cells. Furthermore, it was found that growth medium is superior to PBS as the surrounding solution to enhance the transduction efficiency in some fish cells. In addition, transgene expression can persist for a lengthy period in fish cells, and attained highest level several days later after transduction. This study suggest that the improved recombinant baculovirus can be an excellent gene delivery vector for fish cells and also providing new insights into the transduction of vertebrate cells with baculovirus.
authors.affiliation: College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Fengtao
authors.initials: F
authors.lastname: Huang
authors.forename: Shengbo
authors.initials: S
authors.lastname: Cao
authors.forename: Xiaoxian
authors.initials: X
authors.lastname: Cui
authors.forename: Chuanxi
authors.initials: C
authors.lastname: Xiong
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.forename: Yuanan
authors.initials: Y
authors.lastname: Lu
authors.forename: Weimin
authors.initials: W
authors.lastname: Wang
authors.forename: Jing
authors.initials: J
authors.lastname: Ye
authors.forename: Xueqin
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D003587:Cytomegalovirus / Q000235:genetics; D005399:Fishes; D005434:Flow Cytometry; D022761:Gene Transfer, Horizontal; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014161:Transduction, Genetic / Q000379:methods*
chemical_list: D003470:Culture Media; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins