昆虫病毒多角体蛋白基因的克隆及其在大肠杆菌中的表达  

CLONING AND EXPRESSION OF POLYHEDRIN GENE FROM INSECT VIRUS IN E. COLI CELL

作　　者：齐义鹏[1] 黄永秀 沈英 尹义忠 

机构地区：[1]武汉大学病毒学及分子生物学系

出　　处：《微生物学报》1991年第1期25-31,共7页Acta Microbiologica Sinica

基　　金：国家科学基金;国家教委基金

摘　　要：将含有大尺蠖核型多角体病毒(BsNPV)多角体蛋白基因(ocu)的BamHI-H片段克隆到表达载体pDR 540的BamHI位点,得到能抗高浓度氨苄青霉素(200μg/ml)的两个阳性重组体,其胞外总蛋白比亲本质粒高1.1—1.4倍。在大肠杆菌细胞中,BsNPV ocu基因在原核杂合启动子tac驱动下能高水平的表达,表达量达到0.8和4.7mg/ml。免疫沉淀反应证明,表达蛋白为BsNPV的包涵体蛋白,凝胶过滤法测定的平均分子量为32.1×10~3道尔顿。The BamHI-H fragment containing polyhedrin gene (ocu) of Buzura suppressuria nuclear polyhedrosis virus (BsNPV) was inserted into BamHI site in plasmid pDR540 and two Apr positive recombinants were obtained. Total proteins Produced by clones are more 1.1—1.4 folds than that by parent strain carried plasmid pDR540 in the medium. Expression of ocu protein was 0.8 mg/ml and 0.47 mg/ml. The expression protein was isolated and purified by PAG electrophoresis and gel chromatograph. The molecular weight is 32.1×10~3 dalton.

关 键 词：昆虫病毒 BsNPV ocu 表达蛋白 

分 类 号：S476.13[农业科学—农业昆虫与害虫防治]