肺炎克雷伯菌突变株ΔrcsB的构建及其菌株超粘性与生物膜表型

发布时间：2018-12-17 05:04

【摘要】：目的构建肺炎克雷伯菌rcs B基因缺失的无痕突变株和回补株,通过对突变株的超粘性和生物膜表型进行分析,探讨转录调控子Rcs B对细菌超粘性和生物膜形成能力的影响。方法首先构建突变株Δrcs B,PCR扩增rcs B基因上下游同源臂片段,克隆到质粒p KO3-Km上,将重组质粒导入肺炎克雷伯菌NTUH-K2044中,经同源重组后筛选得到无痕突变株。然后构建回补株,扩增rcs B基因片段,克隆到质粒p BAD33上,将重组质粒导入突变株Δrcs B中,得到回补株。测定野生株、突变株、回补株的超粘性和生物膜形成能力。结果成功构建rcs B基因缺失的无痕突变株Δrcs B和回补株C-Δrcs B。与野生株相比,突变株超粘性和生物膜形成能力均减弱,回补株介于野生株和突变株之间。结论转录调控子Rcs B对肺炎克雷伯菌超粘性和生物膜形成具有正向调控作用。
[Abstract]:Objective to construct the scarless mutant and complement strain of Klebsiella pneumoniae rcs B gene deletion. By analyzing the hyperviscosity and biofilm phenotype of the mutant, the effect of transcription regulator Rcs B on bacterial hyperviscosity and biofilm formation was studied. Methods the upstream and downstream homologous arm fragments of rcs B gene were amplified by 螖 rcs BG-PCR and cloned into plasmid p KO3-Km. The recombinant plasmid was introduced into NTUH-K2044 of Klebsiella pneumoniae. After homologous recombination, trace free mutant was obtained. Then, the complement strain was constructed, the fragment of rcs B gene was amplified and cloned into plasmid p BAD33. The recombinant plasmid was introduced into the mutant 螖 rcs B and the complementary strain was obtained. The hyperviscosity and biofilm formation ability of wild, mutant and compensatory strains were determined. Results the trace free mutant 螖 rcs B and the complement strain C- 螖 rcs B were successfully constructed. Compared with the wild strain, the superviscosity and biofilm formation ability of the mutant were weakened, and the compensatory strain was between the wild and the mutant. Conclusion Transcriptional regulator Rcs B can positively regulate the hyperviscosity and biofilm formation of Klebsiella pneumoniae.
【作者单位】：重庆医科大学公共卫生与管理学院医学与社会发展研究中心健康领域社会风险预测治理协同创新中心;
【基金】：国家自然科学基金(31200064,31071093,31170129)~~
【分类号】：R446.5

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