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【目的】考察炭疽芽胞杆菌中规律成簇的间隔短回文序列(Clustered regularly interspaced short palindromic repeats,CRISPR)位点多态性情况及基于CRISPR位点多态性的分子分型方法是否在炭疽芽胞杆菌分型中适用。【方法】下载NCBI数据库中6株炭疽芽胞杆菌基因组并截取其中CRISPR位点片段序列。根据炭疽芽胞杆菌内CRISPR位点信息,设计相关引物,以193株炭疽芽胞杆菌基因组为模板PCR扩增CRISPR位点片段,测序。本地Blast比对截取序列及测序结果,查看CRISPR位点在炭疽芽胞杆菌中的多态性情况,并比较炭疽芽胞杆菌与蜡样芽胞杆菌和苏云金芽胞杆菌内CRISPR位点情况。【结果】炭疽芽胞杆菌内CRISPR位点不存在多态性。【结论】基于CRISPR位点多态性的分子分型方法不适用于炭疽芽胞杆菌分型,但可以用于区分炭疽芽胞杆菌与蜡样芽胞杆菌和苏云金芽胞杆菌。
【Objective】 To investigate the polymorphism of regularly clustered regularly interspaced short palindromic repeats (CRISPR) loci in Bacillus anthracis and whether the molecular typing method based on CRISPR locus polymorphism is in the anthrax spores Bacillus typing applicable. 【Method】 Six strains of Bacillus anthracis were downloaded from the NCBI database and the CRISPR fragment was intercepted. According to the information of CRISPR locus in B. anthracis, the primers were designed and the CRISPR locus fragment was amplified by PCR from 193 Bacillus anthracis genomes and sequenced. Local Blast compared the intercepted sequences and sequencing results to examine the polymorphism of the CRISPR locus in Bacillus anthracis and compared the CRISPR sites within Bacillus anthracis with Bacillus cereus and Bacillus thuringiensis. 【Result】 There was no polymorphism in CRISPR locus in Bacillus anthracis. 【Conclusion】 The molecular typing method based on polymorphism of CRISPR locus is not suitable for Bacillus anthracis typing, but it can be used to distinguish Bacillus anthracis from Bacillus cereus and Bacillus thuringiensis.