抗虫双价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 Bivalent Gene and Study on Transformation Plant by Laser Microbeam

将苏云金芽孢杆菌伴孢晶体(Bacillus thuringiensis,Bt)蛋白的基因与反枝苋菜凝集素(Amaranthus ret-roflexusagglutinin,ARA)基因构建成双价基因的表达载体,编码一种既能抗鳞翅目害虫,又能抗同翅目蚜虫的杀虫蛋白。把双价基因(Bt-BA)连接到原核表达载体pET28a和植物表达载体pBI121上,经过限制性酶切分析和PCR鉴定,结果表明:含有双价基因的原核和真核重组表达质粒均已构建成功。将该双价基因转入油菜,获得抗性小植株。

Bacillus thuringinesis gene and Amaranthus retroflexus agglutinin gene were ligated into bivalent plant expression vector, which can encode a new lepidopter-resistance and coleopterous insect-resistance protein. The bivalent gene （Bt-BA） were ligated into prokaryotic expression vector （pET28a） and plant expression vector（pBI121 ）. These recombinant plasmids pET-BA and pBI-Bt-BA were confirmed by restriction enzyme and PCR analysis and the results showed that these recombinant plasmids were constructed successfully. The bivalent gene was induced into cole, and the transgenic cole has been obtained.