炭疽芽胞杆菌中CRISPR位点

Clustered regularly interspaced short palindromic repeats (CRISPR) site in Bacillus anthracis

摘要:

摘要:【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。

关键词:关键词:规律成簇间隔短回文重复序列，炭疽芽胞杆菌，分子分型

Abstract:

Abstract:［Objective］ To investigate the polymorphism of clustered regularly interspaced short palindromic repeats (CRISPR) in Bacillu santhracis and the application to molecular typing based on the polymorphism of CRISPR in B.anthracis．［Methods］We downloaded the whole genome sequence of 6 B．anthracis strains and extracted the CRISPR sites．We designed the primers of CRISPR sites and amplified the CRISPR fragments in 193 B．anthracis strains by PCR and sequenced these fragments．In order to reveal the polymorphism of CRISPR in B．anthracis，wealigned all the extracted sequences and sequenced results by local blasting．At the same time，we also analyzed the CRISPR sites in B．cereus and B．thuringiensis．［Results］We did not find any polymorphism of CRISPR in B．anthracis.［Conclusion］The molecular typing approach based on CRISPR polymorphism is not suitable for B．anthracis，but it is possible for us to distinguish B．anthracis from B．cereus and B．thuringiensis.

Key words:Keywords: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)，Bacillus anthracis，molecular typing