Cry6Aa2m基因植物表达载体构建及大豆的遗传转化  

Construction of plant expression vector of cry6Aa2m gene and transformation into Glycine max

作　　者：纪巍[1] 杨丽坤[1] 柏锡[1] 朱延明[1] 才华[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030

出　　处：《东北农业大学学报》2014年第1期59-64,共6页Journal of Northeast Agricultural University

基　　金：国家转基因生物新品种培育重大专项(2009ZX08009-032B)

摘　　要：cry6Aa2杀线虫晶体蛋白基因对线虫有很高的杀虫活性,根据大豆密码子的偏爱性,人工合成杀线虫最小毒力区间cry6Aa2m基因。以pCAMBIA3300为骨架,构建由组成型强启动子E12 CaMV35S调控的cry6Aa2m基因、筛选标记基因为除草剂抗性bar基因的植物表达载体pCBE-cry6,用农杆菌介导法对东农50大豆子叶节进行遗传转化,通过PCR和RT-PCR检测,结果证明cry6Aa2m基因已整合到大豆基因组中并进行转录,获得转cry6Aa2m基因大豆新株系18株,为大豆抗线虫分子育种研究提供材料。Bacillus thuringiensis （Bt） crystal （Cry） proteins are present in the cry6Aa2 intoxicate free-living nematodes. As bacterial codon usage is incompatible with expression in plants, the paper codon-modified cry6Aa2 genes designed by selecting the optimal codon usage for each amino acid based on the soybean genome information. The expression vector pCBE-cry6 was constructed. In the vector, cry6Aa2m gene was driven by promoter E12 CaMV35S. Bar served as selection marker and cry6Aa2m gene was transformed into Dongnong50 via Agrobacterium-mediated method. Total 18 soybean transgenic resistant lines were acquired. PCR and RT-PCR detection indicated that the cry6Aa2m was successfully integrated into soybean genome and expressed. The research materials yielded in this study might be useful on study of resistence to Heterodera glycines.

关 键 词：大豆 cry6Aa2m基因 遗传转化 农杆菌介导 转基因株系 

分 类 号：S572[农业科学—烟草工业]