紫外和EMS诱变选育大肠杆菌赖氨酸营养缺陷型突变株的综合实验设计与实践  

Design and practice of comprehensive experiment of screening lysine mutants by UV and EMS induced mutagenesis

作　　者：颜菲 毕平 张涛 潘皎[2] YAN Fei;BI Ping;ZHANG Tao;PAN Jiao(College of Biomedical Engineering and Technology,Tianjin Medical University,Tianjin 300070,China;College of Life Sciences,Nankai University,Tianjin 300071,China)

机构地区：[1]天津医科大学生物医学工程与技术学院,天津300070 [2]南开大学生命科学学院,天津300071

出　　处：《实验技术与管理》2021年第9期204-209,223,共7页Experimental Technology and Management

基　　金：天津市教委科研计划项目(2019KJ175)。

摘　　要：将科研项目成果应用于本科实验教学,设计了"紫外(UV)和甲基磺酸乙酯(EMS)诱变选育大肠杆菌赖氨酸营养缺陷型菌株(Lys–)"综合实验项目。以大肠杆菌(E.coli DH5a)为出发菌株,通过UV和EMS单一诱变和复合诱变的方法,利用青霉素法淘汰野生型菌株,采用逐个检出法进行初筛、划线对照法进行复检,通过传代培养检测Lys–突变株的遗传稳定性;对青霉素淘汰野生型菌株相关步骤重要参数的确定从原理和方法上进行探讨;通过优化UV、EMS诱变及复合诱变条件,对选育大肠杆菌Lys–突变株的高效诱变方法进行探讨。The results of scientific research projects have been applied to undergraduate experimental teaching, and a comprehensive experimental project of "Ultraviolet(UV) and ethyl methanesulfonate(EMS) induced mutagenesis and screening of Escherichia coli lysine auxotrophic strains(Lys–)" was designed. By taking E.coli DH5 a as the starting strain, and using UV and EMS single mutagenesis and compound mutagenesis methods, the penicillin method is used to eliminate wild-type strains. The one-by-one detection method is adopted for preliminary screening, and streaking control method is used for replication. The genetic stability of the Lys– mutant strain is tested by subculture. The principle and method of determining the important parameters of the steps related to the elimination of wild-type strains of penicillin are discussed. By optimizing the conditions of UV, EMS and compound mutagenesis, the efficient mutagenesis method for screening E.coli Lys– mutant strains is discussed.

关 键 词：实验教学 紫外 甲基磺酸乙酯 赖氨酸营养缺陷型 综合实验设计 

分 类 号：G642.3[文化科学—高等教育学] Q933[文化科学—教育学]