肺炎链球菌转化模型的建立  

Establishment of transformation model of Streptococcus pneumoniae

作　　者：赵清[1] 李南[1] 张雪梅[1] 胥文春[1] 朱兴华[1] 张群[1] 尹一兵[2] 

机构地区：[1]重庆医科大学医学检验系,重庆400016 [2]临床检验诊断学省部共建教育部重点实验室,重庆400016

出　　处：《第四军医大学学报》2008年第9期804-806,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30400376)

摘　　要：目的:建立一种更加稳定、高效的肺炎链球菌实验室转化模型,以便于对该菌的各种目的基因进行重组改造.方法:分别用改进方法和既往方法制备肺炎链球菌感受态细胞,转化外源DNA,获取发生了转化的菌株,通过抗生素培养基培养和PCR鉴定是否发生转化;并计数抗生素筛选平板上的转化菌落,比较两者的转化率,以确定更佳的转化模型.结果:肺炎链球菌334菌株、31203菌株用改进方法的转化率分别为(0.89±0.20)%,(0.71±0.10)%,高于既往方法的转化率[分别为(6.2±1.4)×10-3%,(5.7±1.1)×10-3%],构建了肺炎链球菌感受态缺陷菌株,建立了改进的肺炎链球菌实验室转化模型.结论:改进的转化模型实现了稳定的肺炎链球菌实验室转化,能方便研究者对该菌进行各种遗传操作,为深入研究其致病的分子机制提供了一个技术平台.AIM ： To establish a transformation model of Streptococcus pneumoniae（ S. pn） in laboratory to promote the transformation efficiency and reconstruct its chromosome gene by improving the previous transformation method. METHODS： The competence cells of S. pn were prepared using both the previous and improved methods. After the exotic DNA was transformed under certain cell density, the transformed colonies on antibiotic plate, which was identified by growing in antibiotics culture and PCR, were counted to compare their transformation rate. RESULTS ： The improved transformation model of S. pn in laboratory was established. The transformation rate of S. pn 334 was （0. 89 ± 0.20）% and S. pn 31203 was （0.71 ±0. 10）% under the improved transformation model, which was relatively higher. Using this model, we constructed 4 S. pn competence-defective strains. CONCLUSION： In laboratory, the improved transformation model can raise the transformation rate of S. pn, which induces different S. pn into competence, and provides a research technology for further investigation on pneumococcal pathogenesis.

关 键 词：肺炎链球菌 转化 细菌 感受态 

分 类 号：R378.14[医药卫生—病原生物学]