家蚕转座子表达载体pSVHK1.4的构建  

Construction of Bombyx mori Transposon Expression Vector pSVHK1.4

作　　者：肖巧学[1] 黄亚东[2] 曹阳[1] 刘良式[3] 黄自然[1] 鲁成[4] 余炳球 黄星光 余爱群 

机构地区：[1]华南农业大学蚕业服装系,广州510642 [2]暨南大学医药生物技术研究开发中心 [3]中山大学生命科学学院 [4]农业部蚕桑学重点开放实验室 [5]广东省丝绸集团公司

出　　处：《蚕业科学》2000年第4期228-233,共6页ACTA SERICOLOGICA SINICA

基　　金：广东省科学基金! [A( 970 0 3) ];广东省丝绸 (集团 )公司资助项目

摘　　要：研究利用PCR方法扩增黑腹果蝇 (D .melanogaster)hsp70启动子 ,然后插入到质粒pcDNA3上水母绿色荧光蛋白基因gfp的上游 ,与之顺向拼接构建成含hsp70和gfp的重组质粒pCGH。将家蚕K1 4转座子序列从pUK1 .4亚克隆到真核表达载体pSVL中PvuⅡ位点 ,获得中间载体pSK1 .4。最后将hsp70启动子和gfp报告基因的融合片段从pCGH中亚克隆到质粒pSK1 .4中K1 .4片段上的BglⅡ位点处 ,从而完成对转座子表达载体pSVHK1 .4的构建。In order to construct Bombyx mori transposon expression vector as a tool in research on the transgenic genetics and breeding of silkworm,the hsp70 promoter from D.melanogaster genome was amplified by PCR and cloned into plasmid pcDNA3 GFP containing Green Fluorescent Protein gene( gfp ) firstly to obtain recombinant plasmid pCGH.Secondly,silkworm K1.4 transposable element(1.35kb fragment)from plasmid pUK1.4 was transferred into eukaryon expression vector pSVL to get recombinant plasmid pSK1.4.Lastly,large fragment containing hsp70 promoter and gfp from plasmid pCGH was transferred into Bgl Ⅱ site at the K1.4 transposable element of pSK1.4.The silkworm transposon expression vector pSVHK1.4 has: been contructed succssfully.

关 键 词：家蚕 K1.4转座子 表达载体 构建 

分 类 号：S881[农业科学—特种经济动物饲养]