piggyBac转座子介导的家蚕细胞转基因研究初探  

The Elementary Transgene Research of Silkworm Cells Mediated by Transposon piggyBac

作　　者：周文林[1] 王崇龙[1] 刘波[1] 曹广力[1] 薛仁宇[1] 沈卫德[1] 贡成良[1] 

机构地区：[1]苏州大学生命科学学院,苏州215123

出　　处：《蚕业科学》2007年第1期30-35,共6页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(编号2005CB-121000);国家自然科学基金项目(编号30571404);江苏省研究生培养创新工程项目(编号ZY320506)

摘　　要：为探讨稳定转化家蚕细胞表达外源基因的新方法,以家蚕核型多角体病毒(BmNPV)极早期蛋白基因(ie-1)启动子元件驱动新霉素抗性基因(neor),并克隆至具有绿色荧光蛋白基因(gfp)标记的昆虫转座子载体piggyBac中,构建转基因载体pigA3-IE-Neo。以该载体转染家蚕BmN细胞,用终浓度800μg/mL的遗传霉素(geneticin,G418)筛选3个月,获得了稳定转化的细胞,呈现绿色荧光的细胞数达80%以上。通过PCR鉴定证实了细胞基因组DNA中neor基因和gfp基因的存在。Neomycin resistance gene driven by ie-1 promoter element of Bombyx mori nucleopolyhedrorvirus was cloned into insect transposon vector piggyBac, which contains gfp report gene, to form transgenic vector pigA3-1E-Neo. Then it was used to transfect BmN cell. Screening under G418 at 800 μg/mL final concentration for three months a stable transformation cell strain with over 80% fluorescent cells was obtained. Identification of PCR indicated that neo＇ and gfp genes existed in the genomic DNA of the transgenic cell strain. This study brought forward a new way to express interesting gene by using stable transformation cell.

关 键 词：转基因 家蚕 PIGGYBAC转座子 家蚕细胞 

分 类 号：S881.26[农业科学—特种经济动物饲养]