穿梭质粒匹配pHT254电转枯草芽孢杆菌WB800N方法优化  

Optimization of Shuttle plasmid matching pHT254 for electro-transformation of Bacillus subtilis WB800N

作　　者：吴绍锋 张广献[2] 许晓伍[3] 陈光佩 WU Shao-feng;ZHANG Guang-xian;XU Xiao-wu;CHEN Cuang-pei(Center for Experimental Teaching,School of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Department of Medical Biotechnology,School of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China;Department of Human Anatomy,School of Basic Medical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006,China)

机构地区：[1]广州中医药大学基础医学院实验教学中心,广东广州510006 [2]广州中医药大学基础医学院生物技术系,广东广州510006 [3]广州中医药大学基础医学院解剖学系,广东广州510006

出　　处：《解剖学研究》2023年第2期139-142,共4页Anatomy Research

基　　金：广东省中药局科技基金资助项目(20191097);广东省教学改革项目项目(A1-2601-22-427-003Z060)。

摘　　要：目的 枯草芽孢杆菌虽已广泛应用,但不同质粒转化困难或成本较高,限制了其应用。本研究主要观察了生长培养基、电击缓冲液、复苏培养基中添加如山梨醇、甘露醇、聚乙二醇等不同组合添加剂对枯草芽孢杆菌转化效率及稳定性的影响。方法 用穿梭质粒PHT254电转化枯草芽孢杆菌WB800N,研究生长培养基、电击缓冲液和复苏培养基等因素对转化效率的影响。结果 同一电场压力下高渗体系转化效率较常规体系明显提高且稳定性较好,同一电场压力和高渗体系中使用聚乙二醇PEG6000的电击缓冲液转化效率比使用山梨醇或海藻糖高6倍。结论 高渗电击转化法能够提高转化的稳定性,使用聚乙二醇PEG6000作为缓冲液可以获得更高的电转化效率。Objective Although Bacillus subtilis has been widely used,it is difficult or expensive to transform different plasmids,which limits its application.In this study,the effects of different combinations of additives such as sorbitol,mannitol,and polyethylene glycol on the electroporation efficiency and stability of Bacillus subtilis were observed in growth medium,shock buffer,and recovery medium.Methods The shuttle plasmid PHT254 was used to electro-transform Bacillus subtilis WB800N,and the effects of growth medium,buffer and resuscitation medium on the electroporation efficiency were studied.Results The electroporation efficiency of hypertonic systems under the same electric field pressure was significantly improved and the stability was better than that of conventional systems.In the same electric field pressure and hypertonic system,the conversion efficiency of buffer using polyethylene glycol PEG6000 was six times higher than that of sorbitol or trehalose.Conclusion High osmolarity improved the stability of the electroporation and higher efficiency can be achieved by using peg PEG6000 as buffer.

关 键 词：枯草芽孢杆菌 WB800N PHT254 聚乙二醇 电击转化效率 

分 类 号：Q78[生物学—分子生物学]