抗虫杀菌融合基因表达载体的构建及微束激光转化植物研究  

Construction of Expression Vector with Anti-insect and Antifungal Gene and Study on Transformation Plant by Laser Microbeam

作　　者：张楠[1] 陈正华 刘桂珍 陈凌娜[1] 曲延英[1] 

机构地区：[1]新疆农业大学农学院,新疆乌鲁木齐830052 [2]甘肃亚盛集团博士后科研工作站北京分站,北京100101

出　　处：《激光生物学报》2007年第2期200-207,共8页Acta Laser Biology Sinica

摘　　要：将苏云金芽孢杆菌伴孢晶体蛋白的基因(Bacillus thuringiesis,简称Bt)通过甘氨酸接头(Gly4Ser)3与一种人工合成的抗菌肽(antimicrobial peptides,AMP)基因与相融合,编码一种新的杀虫,并具有抗菌的蛋白。把融合基因(NAMP-Bt)连接到原核表达载体pET-28a和植物表达载体pBI-121上,经过限制性酶切分析和PCR鉴定,结果表明含有融合基因的原核和真核重组表达质粒均已构建成功,并将该融合基因转入烟草,已获得抗性小植株。Antifungal protein gene and Bacillus thuringiesis gene were ligated by the linker of （Gly4Ser）3, which encode a new anti-insect and antifungal protein. The fusion gene （ NAMP -Bt） were ligated into prokaryotic expression vector and plant expression vector. These recombinant plasmids pET-NAMP-Bt and pBI-NAMP-Bt were confirmed by restriction enzyme and PCR analysis and the results showed that these recombinant plasmids were constructed successfully. The fusion gene was induced into tobacco, we had obtained transgenic tobacco.

关 键 词：苏云金芽孢杆菌伴孢晶体蛋白基因 抗菌肽基因 原核表达载体 植物表达载体 限制性酶切分析 PCR鉴定 

分 类 号：Q78[生物学—分子生物学] Q789