猪生长激素基因在杆状病毒载体系统中的表达  

Expression of Porcine Growth Hormone Gene in Baculovirus Vector System

作　　者：欧阳菁[1] 魏平华[2] 杨林[1] 谢明权[2] 杨洁[1] 龙綮新[1] 王珣章[1] 

机构地区：[1]中山大学生物医药中心,生物防治国家重点实验室,广州510275 [2]广东省农业科学院,广州510650

出　　处：《中国病毒学》2000年第3期285-290,共6页Virologica Sinica

基　　金：广东省自然科学基金重点项目!(96 30 0 7)

摘　　要：通过对猪生长激素 (pGH)基因的cDNA进行测序 ,得到 pGHcDNA的全序列 ,并与Seeburg等报道的序列进行了比较和讨论。然后利用具人工合成启动子和多角体蛋白XIV启动子的转移载体质粒 pSXIVVI+ X3/ 4构建出含 pGH基因的重组质粒 pX3/ 4 pGH。将 pX3/ 4 pGH与致死缺失型线性化AcMNPV OCC- DNA共转染Sf9细胞 ,构建出既能形成多角体又能表达 pGH基因的苜蓿丫纹夜蛾核多角体重组病毒AcMNPV pX3/ 4 pGH OCC+ 。感染重组毒株的Hi 5细胞可溶蛋白及其培养上清的SDS PAGE和Westernblot的分析结果表明 ,感染细胞的蛋白电泳带的 2 0 .7kDa处有一条猪生长激素特异带 ,但培养上清中没有。凝胶黑度扫描估测结果显示 pGH蛋白占细胞可溶蛋白的 4 .4 8%。The cDNA sequence encoding porcine growth hormone (pGH) was inserted into the plasmid of pBluescript M13( ). The sequening results showed that a 984 base pair fragment is identical to that reported by Seeburg et al ., with the exception of one different base pair in the ORF. Then, a baculovirus recombinant vector (pX3/4 pGH) with the pGH ORF was constructed and cotransfected Hi 5 cells with the linearized parental virus (AcMNPV OCC -) DNA to produce the recombinant virus AcMNPV pX3/4 pGH OCC +, in which the pGH gene was driven by the synthetic and XIV promoters. The SDS PAGE and Western blot analysis showed that the pGH gene product, 20.7 kDa fusion protein, was obtained. The expressed pGH accumulated up to about 4.48% of the soluble cellular proteins as estimated by CS 930 scanning of the SDS PAGE gel stainning by Coomassie Brilliant Blue R250.

关 键 词：猪生长激素基因 杆状病毒载体系统 表达 

分 类 号：Q786[生物学—分子生物学] S828[农业科学—畜牧学]