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枯草芽胞杆菌紫外线诱变实验方案的优化
引用本文:韦善君,张雪萌,李婧贤,戴景峰,冉坤念,刘立亚,周宜君.枯草芽胞杆菌紫外线诱变实验方案的优化[J].微生物学通报,2016,43(3):695-700.
作者姓名:韦善君  张雪萌  李婧贤  戴景峰  冉坤念  刘立亚  周宜君
作者单位:中央民族大学生命与环境科学学院 北京 100081,中央民族大学生命与环境科学学院 北京 100081,中央民族大学生命与环境科学学院 北京 100081,中央民族大学生命与环境科学学院 北京 100081,中央民族大学生命与环境科学学院 北京 100081,中央民族大学生命与环境科学学院 北京 100081,中央民族大学生命与环境科学学院 北京 100081
基金项目:国家自然科学基金项目(No. 31100507);北京市专业建设与专业综合改革项目(2014);北京市大学生科学研究与创业行动计划项目(No. BEIJ2015110010)
摘    要:【目的】优化枯草芽胞杆菌紫外线诱变实验教学方案,以便学生在实验中获得预期结果。【方法】从菌体培养、活菌浓度估测和辐射参数三方面探讨影响实验结果的因素。【结果】采用液体培养法活化菌体有利于制备均匀的菌悬液,采用比浊法估测初始菌悬液浓度可以指导学生将其稀释至102-103 CFU/m L的浓度;涂布接种后平板表面干燥、培养皿盖内无明显水珠附着,是获得单菌落的关键;在紫外辐射过程中采用固定的菌悬液体积和搅拌速度,可以获得规律性的辐射剂量——致死效应曲线。【结论】优化后的方案实验结果易得、重复性好,可供教学和研究实验参考。

关 键 词:细菌,枯草芽胞杆菌,紫外线诱变,实验教学

Optimized protocol for ultraviolet mutagenesis of Bacillus subtilis
WEI Shan-Jun,ZHANG Xue-Meng,LI Jing-Xian,DAI Jing-Feng,RAN Kun-Nian,LIU Li-Ya and ZHOU Yi-Jun.Optimized protocol for ultraviolet mutagenesis of Bacillus subtilis[J].Microbiology,2016,43(3):695-700.
Authors:WEI Shan-Jun  ZHANG Xue-Meng  LI Jing-Xian  DAI Jing-Feng  RAN Kun-Nian  LIU Li-Ya and ZHOU Yi-Jun
Institution:College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China,College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China,College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China,College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China,College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China,College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China and College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China
Abstract:Objective] To optimize the experiment protocol of ultraviolet (UV) radiation of Bacillus subtilis so that students can get better results in the experiment lesson. Methods] We optimized the protocol by determining the best bacterial cultivation, estimation of viable cells, and UV-radiation parameters. Results] Liquid medium was better than agar slant to obtain a homogeneous suspension of cells. The cell amount of the initial bacterial suspension was estimated efficiently by turbidimetric method, which was convenient to dilute the initial suspension to a concentration of 102?103 CFU/mL. Dry surface covered with dry dish was necessary for the formation of singlet colonies on a bacteria-inoculated plate. During UV exposure, the lethal effect was affected by the adding volume of suspension and mixing speed. Conclusion] The optimized protocol produces repeatable results easily and can be used for teaching or research.
Keywords:Bacterium  Bacillus subtilis  UV mutagenesis  Lab teaching
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