荧光素酶报告基因在转基因蚕中的应用  

Application of Luciferase Report Gene in Transgenic Silkworm

作　　者：陈秀[1] 张峰[2] 赵昀[2] 祁国荣[2] 陆长德[2] 黄君霆[1] 

机构地区：[1]中国农业科学院蚕业研究所,镇江212018 [2]中国科学院上海生物化学研究所,上海200031

出　　处：《高技术通讯》2000年第1期19-22,共4页Chinese High Technology Letters

基　　金：863计划!( 863 1 0 3 1 3 0 2 0 6);国家自然科学基金!395 70 5 5 8资助项目

摘　　要：报道了在针对BmNPV的核酶转基因蚕筛选过程中利用荧光素酶报告基因的检测及鉴定结果。试验采用的质粒为含家蚕核多角体病毒(BmNPV)的即刻早期蛋白基因IE启动子荧光素酶(Luciferase)报告基因———抗NPVIE核酶基因的联合重组质粒pIELucRz。该质粒经限制性内切酶ScaⅠ线性化后,用基因枪法导入家蚕早期受精卵(G0),待G1代测定荧光素酶活性表达较高的蚕的羽化交配产卵,分别于G2、G3和G4代再用BmNPV接种筛选出对NPV抗性较高的蚕继代。其中G4代转基因蚕基因组DNA经PCR测定及Southern杂交分析显示,荧光素酶基因已插入家蚕基因组DNA,并以多拷贝形式存在。这表明荧光素酶基因是转基因蚕研究有效的报告基因之一。The luciferase (Luc) report gene was used as a selective marker in screening anti BmNPV ribzyme transgenic silkworm (Bombyx mori.). The linearized plasmid pIE Luc Rz, in which luciferase gene and ribozyme targeting NPV IE were tademly controlled under BmNPV IE promoter, was introduced into silkworm eggs at early stage by gene gun (G 0). The G 2, G 3, and G 4 offspring of the G 1, which have high luciferase activity, were inoculated by NPV and showed distinct resistance comparing with the non trangenic silkworm. Among the G 4 NPV resistant transgenic silkworms, the PCR analysis and southern blotting results demonstrated that the luceiferase gene was integrated into the silkworm genome with multi copy and it is an effective tool in selection of positive transgenic descendant.

关 键 词：荧光素酶 核酶 转基因蚕 家蚕 抗病育种 

分 类 号：S882[农业科学—特种经济动物饲养]