变形链球菌电击转化条件的优化研究  

An effective strategy to enhance electroporation competence of Streptococcus mutans

作　　者：黄正蔚[1] 刘正[1] 马瑞[1] 唐子圣[1] 朱彩莲[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院,上海200011

出　　处：《中国微生态学杂志》2007年第5期404-405,共2页Chinese Journal of Microecology

基　　金：国家自然科学基金(编号30400497);上海市科委重点科研基金(编号04JC14066)资助项目

摘　　要：目的为增强对变形链球菌的电击转化效率,探索常用的胞壁弱化剂甘氨酸在电击转化中的加入模式。方法以氨苄青霉素抗性的pGL3 basic质粒作为外源DNA,通过电击转化导入变形链球菌参考株UA159内,并在选择性培养基上筛选阳性转化克隆,以优化筛选出最佳的甘氨酸加入浓度与模式,同时比较了不同电击方案转化效率的差异。结果在变形链球菌对数生长期后加入终浓度为10%的甘氨酸可有效地增强电击转化的效率;而不同的电转电压的选择对于转化效率的影响,在本实验中差异未见显著性。结论研究证实了甘氨酸作为胞壁弱化剂可增强对变形链球菌的转化效率,并优化了对变形链球菌的电击转化方案。Objective To optimize the transform efficiency, compare different electrophoration competence methods. Methods pGL3 basic plasmids with Ampicillin resistance was used as foreign DNA to be introduced into Streptococcus mutans cells. Then positive transformants were counted on selective agar. Different glycine concentration together with different electroporation voltage were compared to optimize the transformation methods. Results The final concentration of 10% glycine could effectively enhance transformation efficiency,while different electroporation voltages showed no significance in this study. Conclusions As a cell-wall weakening agent,glycine can be used to enhance electroporation competence. This study also optimized the transform protocols to Streptococcus mutans.

关 键 词：变形链球菌 电击转化 甘氨酸 

分 类 号：R378.12[医药卫生—病原生物学]