摘要
为分离鉴定棉田转Cry1Ac基因棉的Cry1Ac蛋白土壤降解细菌及测定其降解能力。采用蛋白质凝胶电泳(SDSPAGE)、酶联免疫法(ELISA)法和常规细菌培养与鉴定技术,对第一师12团棉田土壤中Cry1Ac蛋白降解菌进行分离、纯化、形态生理生化观察以及降解功能测定。结果表明:从棉田土壤分离一株具有降解Cry1Ac蛋白能力的土壤细菌,命名为12T-103菌株,细菌形态学、生理生化特征和16S r DNA序列鉴定为枯草芽孢杆菌(Bacillus subtilis),该菌株对Cry1Ac蛋白的降解效果较为明显,在1h时Cry1Ac蛋白含量由15. 95μg·L-1降至0. 68μg·L-1,其降解程度达极显著水平(p <0. 01),其降解率为92. 26%。结论:分离于转基因棉田的12T-103菌株为枯草芽孢杆菌(Bacillus subtilis),该菌株对Cry1Ac蛋白的降解能力较强。
For isolation and identification of Cry1Ac protein degrading bacterial strain and its degradation ability from the soil of planting Bt cotton in the farm 12,first division of Xinjiang production and construction corps,the Cry1Ac protein degrading bacterial strain and its bacterial form,physiology and biochemistry in the soil were isolated,purified and detected with SDS-PAGE,enzyme linked immunoassay(ELISA)and conventional bacterial culture and identification technologies.The results showed that one Cry1Ac protein degrading bacterial strain was isolated and named as 12T-103,and the 12T-103 was identified as Bacillus subtilis according to the phenotype observation,characteristics of physiology and biochemistry and 16S rDNA analysis,which had great significant effect on the degradation of Cry1Ac protein from15.95μg·L-1to0.68μg·L-1in1h(p<0.01),and the degradation rate of Cry1Ac protein was92.26%.It indicated that the12T-103from the Bt cotton planting soil was Bacillus subtilis,and had strong degradation ability on Cry1Ac protein.
作者
肖海兵
董震宇
董红强
符娜
尚北晨
杨明禄
Xiao Haibing;Dong Zhenyu;Dong Hongqiang;Fu Na;Shang Beichen;Yang Minglu(College of Plant Science,Tarim University,Alar,Xinjiang 843300)
出处
《塔里木大学学报》
2018年第4期59-63,共5页
Journal of Tarim University
基金
塔里木大学校长基金青年项目"南疆棉田Cry1Ac毒蛋白土壤细菌降解效能研究"(TDZKQN201601)
塔里木大学大学生创新创业训练计划项目"12T-103菌株对Cry1Ac毒蛋白降解特征分析"(107572017052)
