摘要
研究了口蹄疫病毒(FMDV)VP31基因在昆虫细胞中的表达.结果表明:利用重组杆状病毒在昆虫细胞中成功表达了AsiaⅠ型口蹄疫病毒(FMDV)结构蛋白VP31基因,目的基因经亚克隆插入到含有一段蜂毒溶血肽序列的转移载体pMelBac B中,构建出穿梭质粒pMel-VP31.将含有目的基因的穿梭载体与线性化的杆状病毒骨架共转染昆虫Sf9细胞,通过噬斑筛选和PCR鉴定,获得了含有目的基因的重组杆状病毒.重组病毒经扩增后感染Sf9细胞,用Western-blot和间接夹心ELISA检测,证实VP31蛋白获得了表达.
The structural protein gene VP31 of Asia I foot-and-mouth disease virus（FMDV） was expressed in the insect cells.VP31 gene was subcloned into a shuttle vector of pMelBac-B with a melittin secretion signal sequence to get recombinant plasmid of pMel-VP31.The recombinant shuttle vector was cotransfected with linearized Bac-N-BlueTM DNA into Sf9 insect cell to get recombinant baculovirus by plaque screening and PCR identification.The Sf9 cells were infected with recombinant baculoviruses after primary amplification.The cells were harvested after 72 h of infection with recombinant baculovirus.The products were analyzed with Western-blot and indirect sandwich ELISA,and the results showed that the VP31 gene were successfully expressed in insect cells.This work made a valuable investigation for development of new-type of FMDV antigen for vaccine and diagnosis purpose.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2009年第5期16-19,54,共5页
Journal of Gansu Agricultural University
基金
国家“863”项目(2006AA10A2041)
