用重组痘苗病毒表达pGHcDNA的研究  

Expression of pGH cDNA Gene in H143 Cells Using Vaccinia Virus

作　　者：路丹红[1] 朱宝利[1] 周顺伍[1] 齐顺章[1] 

机构地区：[1]北京农业大学动物生物化学教研室

出　　处：《生物化学杂志》1995年第5期504-509,共6页

基　　金：博士点基金

摘　　要：构建了含有ｐＧＨｃＤＮＡ的重组痘苗病毒，用ＥＬＩＳＡ证明该重组病毒在被感染的ｈ１４３细胞中，可表达出猪生长激素并将之分泌到培养基中，表达量约为１．０５μｇ／１０￣６细胞（２４ｈ）。用定位免疫化学法进一步证明该病毒可感染小鼠并在小鼠体内表达ｐＧＨｃＤＮＡ。同时还构建了含双拷贝ｐＧＨｃＤＮＡ的重组痘苗病毒，并证明其ｐＧＨ表达量比单拷贝重组病毒有明显提高，约为１．５０μｇ／１０￣６细胞（２４ｈ）。The porcine growth hormone (pGH) cDNA gene was inserted downstream of the vaccinia virus P_7.5 promoter of plasmid PSC11. Following homologous recombination in vivo,recombinant vaccinia virus was obtained, which carrying the pGH cDNA gene. By infecting h143 cells with this virus, the pGH cDNA gene was expressed under the control of P_7.5 promoter. As estimated by ELISA, the expression efficiency was approximately 1. 05 μg/10￣6 cells in 24 h. The recombinant vaccinia virus was also used to infect the mouse, and the expression product was detected by in situ immuno-chemical analysis. Moreover, recombinant virus with two copies of pGH cDNA was constructed and used to infect h143, where the expression efficiency was approximately 1.50μg/10￣6 cells in 24h, evidently higher than in the infected h143 cells with recombinant virus carrying a single copy of pGH cDNA gene.

关 键 词：猪生长激素 生长激素 痘苗病毒 基因表达 CDNA 

分 类 号：Q575.11[生物学—生物化学]