枯草芽孢杆菌菌株ZT4-2电击转化体系的建立  

Establishment of the electro-transformation system of Bacillus subtilis ZT4-2

作　　者：蔡露 秦艳秋 高坦坦 王旭东 任争光[1] 赵晓燕[1] CAI Lu;QIN Yanqiu;GAO Tantan;WANG Xudong;REN Zhengguang;ZHAO Xiaoyan(College of Bioscience and Resource Environment/Key Laboratory of Urban Agriculture(North China),Ministry of Agriculture and Rural Affairs,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院生物与资源环境学院/农业农村部华北都市农业重点实验室,北京102206

出　　处：《北京农学院学报》2022年第1期32-36,共5页Journal of Beijing University of Agriculture

基　　金：北京市教委科技计划面上项目(KM201910020009)。

摘　　要：【目的】建立枯草芽孢杆菌(Bacillus subtilis)菌株ZT4-2的高效电击转化体系。【方法】带有荧光蛋白标记的穿梭质粒pGFP78,通过电击转化法将其转入枯草芽孢杆菌菌株ZT4-2。【结果】菌株培养阶段、感受态细胞浓度、感受态细胞量、质粒体积量、转化电压等条件的不同,会对电击转化效率造成影响。其中试验条件:制备感受态细胞的菌株培养阶段的OD_(600 nm)值=1.0,重悬菌体时每100mL起始菌液加入600μL的40%PEG6000;电击转化体系中,感受态细胞量200μL,质粒体积10μL,转化电压2.5kV。最高转化效率可达6823.67CFU/μg。【结论】建立了枯草芽孢杆菌菌株ZT4-2的电击转化体系,电击转化效率较高。【Objective】To establish a high-efficiency electro-transformation system for Bacillus subtilis strain ZT4-2.【Methods】The shuttle plasmids pGFP78was transferred into Bacillus subtilis ZT4-2by electro-transformation.【Results】The results showed that electro-transformation efficiency was affected by the bacteria culture stage,competent cell concentration,competent cell amount,plasmid volume,electric shock voltage,etc.The optimal conditions were as follows:The OD_(600 nm)value was 1.0in the culture stage of the strain to prepare the competent cells.Addition of 600μL 40%PEG6000to 100mL initial bacterial solution was needed when the cells re-suspension was prepared.In the electro-transformation system,the amount of competent cells was 200μL,the plasmid volume was 10μL,and the electric shock voltage was 2.5kV.At this condition,the electro-transformation efficiency was the highest,up to 6823.67CFU/μg.【Conclusion】An electro-transformation system of Bacillus subtilis ZT4-2was established,the electro-transformation efficiency was high and may provide a foundation for the molecular genetic research of Bacillus subtilis ZT4-2.

关 键 词：枯草芽孢杆菌菌株ZT4-2 电击转化 聚乙二醇 体系建立 

分 类 号：O657.33[理学—分析化学]