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牛福星,于晶晶,汤宏赤,刘金平,滕昆,韦宇拓.枯草芽孢杆菌突变株IA857高效转化方法的研究[J].广西科学,2014,21(2):119-123. [点击复制]
- NIU Fu-xing,YU Jing-jing,TANG Hong-chi,LIU Jin-ping,TENG Kun,WEI Yu-tuo.Studying on the Efficient Transformation Method for Mutant Bacillus Subtilis IA857[J].Guangxi Sciences,2014,21(2):119-123. [点击复制]
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| 枯草芽孢杆菌突变株IA857高效转化方法的研究 |
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牛福星1,2, 于晶晶1,2, 汤宏赤1,2, 刘金平1,2, 滕昆1,2, 韦宇拓1,2
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| (1.广西大学生命科学与技术学院, 广西南宁 530005;2.亚热带农业生物资源保护与利用国家重点实验室, 广西南宁 530005) |
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| 摘要: |
| [目的]研究比较枯草芽孢杆菌突变株IA857高效转化的最优方法。[方法]以质粒pHCMC04(大小为8089bp)转化突变型枯草芽孢杆菌IA857,分别用电转化法、原生质体法、电击法诱导的原生质体法、Spizizen低盐环境感受态转化法(简称S法)、低盐环境形成饥饿感受态法及其改良方案在其它突变菌株的试验方法(简称C法)进行实验,通过比较优化得出最优转化方法。[结果]采用S法转化枯草芽孢杆菌突变株IA857,质粒加入量为70ng时,转化后直接摇床培养1h转化率最高达到1.2×104 CFU/μg DNA,可以满足枯草芽孢杆菌高效转化的要求。[结论]得出了枯草芽孢杆菌突变株IA857最优转化方法并提出转化枯草芽孢杆菌的基本框架。 |
| 关键词: 枯草芽孢杆菌 转化 电击法 原生质体法 低盐环境 感受态 |
| DOI： |
| 投稿时间：2014-02-18修订日期：2014-03-04 |
| 基金项目:广西科学研究与技术开发计划(No.11107008-3),广西研究生教育创新计划项目资助。 |
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| Studying on the Efficient Transformation Method for Mutant Bacillus Subtilis IA857 |
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NIU Fu-xing1,2, YU Jing-jing1,2, TANG Hong-chi1,2, LIU Jin-ping1,2, TENG Kun1,2, WEI Yu-tuo1,2
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| (1.College of Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;2.State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China) |
| Abstract: |
| [Objective] Studying and comparing efficient transformation methods for mutant B.subtilis IA857.[Methods] The plasmid of pHCMC04 (8089bp) has been transformated into B. subtilis IA857, and many methods, just like electroporation, protoplast method, protoplast induced by electroporation, the method forming hunger feeling competent in low salt environment (short S method) and it's improved method used in other strains (short C method), were compared and optimized.[Results] The optimization of conversion rate was conducted by means of S method.When the mutant B.subtilis IA857 was transformed with 70 ng plasmid and directly shaker cultured for 1h, the highest transformation efficiency reached 1.2×104CFU/μg DNA, which meets the requirement for efficient transformation.[Conclusion] This study provides an optimal transformation method and puts forward the basic framework of transformation of Bacillus subtilis. |
| Key words: B.subtilis transformation electro-poration protoplast method low salt environment competence |
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