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目的:研究霍乱弧菌中TLC基因簇的分子组成特征和类型检测方法的建立。方法:针对TLC基因簇内部的特征性基因设计结构相关引物和探针,利用序列比对、PCR和核酸杂交等分子生物学方法,检测并验证TLC在菌株内的基因结构和排列方式,确定分布类型。结果:pTLC与TLC基因簇之间序列有变异,TLC多整合于染色体中;建立了TLC基因簇的组成类型检测方法,并发现3种不同的结构排列方式。结论:TLC基因簇类型检测方法的建立及变异性的研究,利于深入揭示其在霍乱进化和毒力转移等过程中的影响作用。
OBJECTIVE: To study the molecular characterization and type detection of TLC gene cluster in Vibrio cholerae. Methods: Structurally related primers and probes were designed according to the characteristic genes in TLC gene cluster. The molecular structure and alignment of TLC in strains were determined and verified by molecular biology methods such as sequence alignment, PCR and nucleic acid hybridization Types of. Results: The sequences of pTLC and TLC gene cluster varied, and TLC was more integrated in the chromosome. The method of detecting the composition type of TLC gene cluster was established, and three different structures were found. Conclusion: The establishment of TLC genotyping method and the study of its variability will be helpful to reveal its influence on evolution and virulence of cholera.