苏云金芽胞杆菌标记重组菌株的构建与杀虫基因水平转移

利用SOE法将构建的绿色荧光蛋白基因gfp和苏云金芽胞杆菌的杀虫晶体蛋白基因cry1Ac10的嵌合基因克隆到穿梭载体pAD4412上获得重组质粒pBMBZGC10，再通过电转化法导入苏云金芽胞杆菌无质粒突变株CryB中获得重组菌株CryB(pBMBZGC10).将重组菌株CryB(pBMBZGC10)的发酵液按30、60和90 ml共3个浓度梯度，菌数约为10.7～10.8·ml^-1，分次喷洒供试植株小白菜、蕹菜和番茄,结果表明,cry1Ac10基因没有向供试土壤细菌、真菌和放线菌转移，也未在供试植物根、茎和叶中检测到该基因.

Construction of Bacillus thuringiensis labeled recombinant strain and horizontal transfer of its cry1AC10 gene

A recombinant plasmid pBMBZGC10 was obtained by the ligation of gfp-cry1Ac10 fusion gene and vector plasmid pAD4412,which was then introduced by gene pulser into acrystalliferous strain CryB,and a recombinant strain CryB(pBMBZGC10)was obtained.Different fermentative solutions of recombinant strain were used for multi-spraying on Brassica pekinesis,Ipomoea aquatica and Lycopersicon esculentum leaves.The results of fluorescent detection and PCR amplification revealed that cry1Ac10 gene did not transfer into indigenous bacteria,actinomyces and fungi in test soil,and could not be detected in roots,stems and leaves of test plants.