肺炎链球菌转化模型的建立与优化  

The Establishment and Optimization of Transformation Model of Streptococcus pneumoniae

作　　者：孟江萍[1] 尹一兵[1] 张雪梅[1] 蓝锴[1] 黄远帅[1] 涂植光[1] 

机构地区：[1]重庆医科大学医学检验系重庆市临床检验诊断学重点实验室,重庆400016

出　　处：《微生物学杂志》2006年第1期10-13,共4页Journal of Microbiology

基　　金：国家自然科学基金(30400376)

摘　　要：建立肺炎链球菌转化模型，优化转化体系，提高转化率，以便于进一步研究其致病的分子机制。制备肺炎链球菌感受态。首先在不同菌密度下转化外源DNA，计数抗生素筛选平板上的转化菌落，比较其转化率，确定转化的最适菌密度；然后在此菌密度下比较CSP诱导不同时相的转化率，同时用RT-PCR检测感受态调控基因comE的表这。对所用血清3型菌株而言转化的最适菌密度在OD550=0．09～0．10之间；CSP-2诱导10min后转化率最高，可达（15．6±3）％；comE的表达也在CSP-2诱导10min后达到最高。在实验室条件下。肺炎链球菌转化受多种因素的影响，必需控制好各种因素。选择最优条件才能获得稳定、高效的转化。In order to study the molecular pathogenesis mechanism of Streptococcus pneumoniae （SP） further, its transformation model was established to optimize the transformation system and to increase the transformation efficiency; competence cells of SP 31203 were prepared. First exotic DNA was transformed under different cell density then the transformed colonies on antibiotic screening plate was counted to compare their transformation rate to ascertain the most suitable cell density for the transformation. Under this cell density compare the transformation rate at different time and induced by CSP-2. And at the same time, the expression of regulator and control gene comE of competence was tested with RT-PCR. To the strains of type 3 used in the experiment the most suitable cell density was at OD550 = 0.09-0. 101 and ten minutes alter the transformation rate induced by CSP-2 was the highest and could reach up to （15.6 ± 3） % ; and the expression of come also reached to its peak ten minutes after the induction. Under the lab conditions, the transformation of SP was affected by many factors, therefore controlling them well and select the optimum conditions stable and high efficiency transformation rate can be obtained.

关 键 词：肺炎链球菌 感受态 转化 

分 类 号：R378.14[医药卫生—病原生物学]