重组家蚕病毒表达系统转移载体的构建和β─半乳糖苷酶的表达  

CONSTRUCTION OF A TRANSFER VECTOR pUBM-4 FOR GENE EXPRESSION IN SILKWORM AND EXPRESSIONOF β-GALACTOSIDASE GENE

作　　者：张颖[1] 吴祥甫[1] 

机构地区：[1]中国科学院上海生物化学研究所

出　　处：《病毒学报》1994年第3期251-256,共6页Chinese Journal of Virology

摘　　要：本文从中国农科院蚕业研究所提供的我国家蚕核多角体病毒镇江株中获得了多角体蛋白的强启动子，用此启动子构建了家蚕病毒表达系统的转移载体。外源基因能在此启动子控制下在家蚕细胞和虫体中进行高效表达。用此载体我们首先成功地在家蚕虫体中高效表达了β－半乳糖苷酶，表达量达到５８０μｇ／条蚕，从而证实我们构建的载体是可靠的、有效的，可用于家蚕重组病毒表达外源基因的研究。e have contructed an universal transfer vector pUBM-4 for gene expression in silkworm.In this vector,the promoter is a strong one obtained from the polyhedrin ofBmNPV,Zhengjiang strain;5’flank sequence and 3’flank sequence of polyhedrin serve asthe homologous regions for in vivo recombination.ATG,the translation site of polyhedrin wasmutated to EcoRV site by PCR.The foreign egnes can be inserted into EcoRV site orBamHI site in this vector and expressed under the control of the polyhedrin promoter inBm-cells and larvae.We have sucessfully expressed β-galactosidase gene with this vector insilkworm.The maximum level of expression is 580μg per larva,demonstrating that the transfervector pUBM-4 is reliable and useful.

关 键 词：家蚕 核型 多角体病毒 半乳糖苷酶 

分 类 号：S884.5[农业科学—特种经济动物饲养]