宿主域扩大的重组救活昆虫杆状病毒表达载体的构建及外源基因的表达  

Construction of the Host Range Expanded Recombinant-Rescure Inscet Baculovirus Expression Vector and the Expression of Foreign Gene

作　　者：易咏竹[1] 陈寅[1] 张志芳[1] 何家禄[1] 秦俭[2] 

机构地区：[1]农业部家蚕生物技术重点开放实验室,中国农业科学院蚕业研究所,镇江212018 [2]四川华神集团

出　　处：《蚕业科学》2002年第4期304-307,共4页ACTA SERICOLOGICA SINICA

摘　　要：通过克隆的家蚕核多角体病毒解旋酶基因DNA与重组救活可线性化的苜蓿尺蠖核多角体病毒基因工程载体病毒 (BacPAK6 )DNA在昆虫细胞中发生重组 ,经累代筛选获得了既可感染家蚕又可感染秋粘虫细胞Sf 2 1的宿主域扩大的昆虫杆状病毒表达载体 (HyBacPAK6 )。HyBacPAK6DNA经Bsu36Ⅰ酶切后与含植酸酶基因的转移载体pVL1393 phy在家蚕细胞中重组后 ,通过蓝白斑筛选发现重组率可达 90 %以上。然而以杂交病毒为载体的外源基因表达量仅为以BmNPV为载体病毒的表达量的 2 0 %左右。分析认为HyBacPAK6可用于表达一些对蚕体有害的外源基因。The helicase gene DNA of Bombyx mori Nucleopolyhedrosis Virus (BmNPV) and the viral expression vector BacPAK6 of the Autographa californica Nucleopolyhedrosis Virus (AcNPV),Recombinant-Rescure with linerized,were recombined within insect cells.The host range expanded insect baculovirus expression vector was constructed via seven round selection,which could infect Bombyx mori and Sf-21 cell lines.After digested by Bsu36Ⅰ,the linerized viral DNA and the transferring vector pVL1393-phy which contains the gene of phytase were co-transfected into Bm-5 cell lines.It was found that the recombining rate was above 90% through blue-white plaque selection.But the expression level with hybrid viral vector is only 20% compared to BmNPV viral expression vector,and the HyBacPAK6 may be useful to express the foreign gene which is harmful to silkworm larvae.

关 键 词：表达载体 宿主域 外源基因表达 家蚕 昆虫杆状病毒表达系统 HyBacPAK6 

分 类 号：S884[农业科学—特种经济动物饲养]