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BmNPV和AcNPV扩大寄主域杂交重组病毒表达载体的构建和改进 被引量:15

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摘要 苜蓿银纹夜蛾核型多角体病毒(Autographacalifornicanuclearpolyhedrosisvirus,简称AcNPV)和家蚕核型多角体病毒(Bombyxmorinuclearpolyhedrosisvirus,简称BmNPV)表达系统是目前最主要的两类昆虫杆状病毒表达系统.两者各具特点,前者寄主昆虫个体较小,但寄主范围广、适合较大规模的细胞悬浮培养生产体系;而后者虽然寄主专一,但以个体较大、易饲养的家蚕为寄主,具有表达量高、特别适合产业化水平开发的要求.为了利用AcNPV和BmNPV各自的优势,扩大昆虫杆状病毒的寄主范围,以BmNPV为基础,利用决定病毒寄主范围的DNA解旋酶基因及同源重组原理,构建了介于BmNPV和AcNPV间的杂交重组病毒(hybridbaculovirusofAcNPVandBmNPV,简称HyNPV).这样构建的杂交重组病毒具有AcNPV和BmNPV的双重优点,既能感染所有AcNPV的寄主,又可以在家蚕中表达.以人碱性成纤维细胞成长因子基因作为应用实例,克隆构建了这样的一种重组杂交病毒,可在Sf21,Tn-5,BmN培养细胞及家蚕个体中穿梭表达.为了减少表达后重组蛋白的降解,利用基因敲除法删除了杆状病毒本身的一种主要蛋白酶-半胱氨酸蛋白酶基因,提高了重组蛋白的稳定性和表达效率.
出处 《中国科学(C辑)》 CSCD 北大核心 2004年第2期156-164,共9页 Science in China(Series C)
基金 国家农业部(2003-Z50) 浙江省科技厅重点国际合作项目(2002C24003)
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