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

应用生态学报 ›› 2005, Vol. 16 ›› Issue (1): 142-146.

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

周琴, 孙明, 李林, 杨在清, 喻子牛

华中农业大学农业微生物学国家重点实验室, 武汉, 430070

收稿日期:2003-11-18 修回日期:2004-03-02 出版日期:2005-01-15 发布日期:2005-01-15
通讯作者: 喻子牛
基金资助:
国家自然科学基金项目(30170032)、国家高技术发展计划"863"项目(2004AA214092,2003AA223081)和农业微生物学国家重点实验室基金资助项目.

Construction of Bacillus thuringiensis labeled recombinant strain and horizontal transfer of its cry1A_C10 gene

ZHOU Qin, SUN Ming, LI Lin, YANG Zaiqing, YU Ziniu

State Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China

Received:2003-11-18 Revised:2004-03-02 Online:2005-01-15 Published:2005-01-15

摘要/Abstract

摘要： 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90ml共3个浓度梯度,菌数约为107～108·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.

关键词: 苏云金芽胞杆菌, 绿色荧光蛋白基因(gfp), 杀虫晶体蛋白基因, 水平转移

Abstract: 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.

Key words: Bacillus thuringiensis, Green fluorescent gene gfp, Insecticide crystal gene protein gene cry, Horizontal gene transfer

中图分类号:

Q784

周琴, 孙明, 李林, 杨在清, 喻子牛. 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[J]. 应用生态学报, 2005, 16(1): 142-146.

ZHOU Qin, SUN Ming, LI Lin, YANG Zaiqing, YU Ziniu . Construction of Bacillus thuringiensis labeled recombinant strain and horizontal transfer of its cry1A_C10 gene [J]. Chinese Journal of Applied Ecology, 2005, 16(1): 142-146.

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