家蚕Polh^+ Bac-to-Bac杆状病毒表达系统的构建  

Construction of a BmNPV Polyhedrin-plus Bac-to-Bac System for the Silkworm,Bombyx mori

作　　者：曹翠平[1] 兰丽盼[1] 姚慧鹏[1] 何芳青[1] 郭爱芹[1] 吴小锋[1] 

机构地区：[1]浙江大学动物科学学院,杭州310029

出　　处：《蚕业科学》2008年第2期237-243,共7页ACTA SERICOLOGICA SINICA

基　　金：国家高技术研究发展计划"863"项目(编号2007AA10Z159);浙江省科技厅重点国际合作项目(编号2006C24015);教育部新世纪人才计划项目(编号NCET-06-0524)

摘　　要：为利用杆状病毒的强启动子——多角体启动子而构建的重组杆状病毒一般是多角体缺失型(polh-)病毒。为了解决家蚕生物反应器规模化生产中病毒必需经皮接种而效率低下的问题,在建立家蚕Bac-to-Bac快速表达系统的基础上,构建了能形成多角体的家蚕Polh+ Bac-to-Bac表达系统。通过该系统能够产生表达外源目的基因的重组病毒,获得的该重组病毒能在培养细胞内表达,也能通过经口添食感染表达。利用EGFP报告基因分析了该系统的表达效果,构建的重组病毒不仅有效表达了EGFP蛋白,还在细胞核中形成了大量多角体。该系统较好解决了重组病毒必需经皮接种感染的缺陷,提高了生产效率,拓宽了杆状病毒在生物杀虫剂、基因治疗等领域的应用前景。Usually the recombinant baculovirus was generated through replacing the foreign gene into the polyhedrin locus resulting in a polyhedrin-negative virus. Such polh-negative baculovirus can infect insect cell lines well and insect also through subcutaneous injection, but is less infective to silkworm through per oral in- gestion. In order to improve the inoculation efficiency, a polyhedrin-plus Bac-to-Bac expression system for application in silkworm was constructed in this paper. Using this new system, the foreign gene and polyhedrin were both expressed at higher level and large amount of polyhedra were formed. Thereby the recombinant baculovirus can infect silkworm larva by oral administration. This new system also widen the application of baculovirus in the field of biological insectlcide, gene therapy and other aspects.

关 键 词：家蚕 杆状病毒表达系统 多角体 经口感染 

分 类 号：Q78[生物学—分子生物学]