南方医科大学学报

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (09): 1135-1139.

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B型链球菌数量感应通路测定及LuxS缺失突变株的构建

欧阳谦1, 马文丽1, 刘翠华1, 石嵘1, 郑文岭2   

  1. 1. 南方医科大学基因工程研究所, 广东, 广州, 510515;
    2. 华南基因组中心, 广东, 广州, 510800
  • 出版日期:2005-09-20 发布日期:2005-09-20
  • 基金资助:
    收稿日期:2005-3-15。
    作者简介:欧阳谦,男,博士研究生,E-mail:oyq@fimmu.com
    通讯作者:马文丽,电话:020-61648210,020-61647755,E-mail:wen-li@fim m u.edu.cn.

Detection of quorum-sensing pathway and construction of LuxS gene deletion mutants of group B Streptococcus

OU Yang-qian1, MA Wen-li1, LIU Cui-hua1, SHI Rong1, ZHENG Wen-ling2   

  1. 1. 南方医科大学基因工程研究所, 广东, 广州, 510515;
    2. 华南基因组中心, 广东, 广州, 510800
  • Online:2005-09-20 Published:2005-09-20

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摘要： 目的 对GBS中可能存在的AI-2数量感应通路进行测定,构建数量感应相关分子LuxS缺失突变株并对其基因型进行确认。方法 培养GBS至不同D(λ)值,利用V.harveyi BB170作为报告菌株,对GBS诱导生物发光的能力进行测定,然后寻找GBS中LuxS同源基因,通过同源重组法在GBS中构建LuxS缺失突变株,Southern杂交分析确证。结果 GBS中的某种分泌成份可在报告菌株中诱导生物发光活性,提示GBS中存在AI-2数量感应通路,在GBS中发现了LuxS同源基因并成功地构建了LuxS缺失突变株。结论 本研究为探索LuxS在GBS中AI-2数量感应通路中的重要性及GBS毒力研究建立了细菌模型。

Abstract: Objective To detect the possible presence of AI-2 quorum-sensing pathway and construct group B Streptococcus (GBS) mutants with deletion of LuxS gene related to quorum-sensing pathway. Method V. harveyi BB170 was employed as the reporter strain to detect AI-2 pathway in GBS, and identification of LuxS homologous gene in GBS type V strain 2603 was performed by software-based analysis. LuxS gene deletion mutant ΔLusX was then constructed in GBS by means of allelic exchange and verified by Southern hybridization analysis. Results A component in the secretions of GBS could induce bioluminescence activity in the reporter strain, suggesting the presence of AI-2 quorum-sensing pathway in GBS. luxS homologous gene was detected in GBS and LuxS gene deletion mutants was successfully constructed in GBS Ia 515 and V 2603 strains. Conclusion This study establishes a bacterial model for studying the role of LuxS molecule in AI-2 quorum-sensing pathway in GBS and provides new insights into virulence regulation mechanism of GBS.

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