保加利亚乳杆菌CRISPR位点的序列分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0952

摘要/Abstract

摘要： 乳酸菌基因组的CRISPR序列是乳酸菌识别并抵御噬菌体侵染的重要元件,其与cas蛋白共同构成了乳酸菌的获得性免疫系统。而目前对乳酸菌CRISPR序列信息研究较少。因此本文对8株不同来源的保加利亚乳杆菌的CRISPR序列进行了克隆测序,对其重复序列进行了遗传进化分析并预测了二级结构,同时进行了间隔序列的同源性分析。结果显示：8株保加利亚乳杆菌均含有长度不一的CRISPR区,最长的CRISPR序列片段长度为1 820 bp,含有30条间隔序列,最小的CRISPR片段仅有408 bp,含5个间隔序列。经分析发现CRISPR重复序列的二级结构预测有两类不同的结构,一类以“环”为主的典型RNA二级结构,一类以“茎”为主,前者剪切后具有典型crRNA结构,而后者的功能还有待进一步研究。通过分析保加利亚乳杆菌的CRISPR序列结构,可为保加利亚乳杆菌抗噬菌体的分子机制奠定基础,也对乳品工业筛选抗噬菌体的发酵剂菌株具有指导意义。

关键词: 保加利亚乳杆菌, 同源性, CRISPR, 间隔序列, 二级结构

Abstract: CRISPR（Clustered regularly interspaced short palindromic repeats）sequence in the genome of Lactobacillus is its important part to recognize and resist phage,and CRISPR and cas protein constitute the acquired immune system. Currently there are few studies on the CRISPR sequence of Lactobacillus. In this study,clone sequencing was performed in the CRISPR of 8 strains of L. bulgaricus from different sources,and then the evolutionary genetics analysis to the repetitive sequence was also conducted,and the secondary structure was predicted;moreover the homology analysis in the spacer sequence was also carried out. Results showed that the lengths of CRISPR in 8 different strains of L. bulgaricus varied,among which the longest one was 1820 bp with 30 spacer sequences,and the shortest one was 408 bp with 5 spacer sequences. Two different structures were found in the prediction on the secondary structure of CRISPR repetitive sequence：one was the typical RNA mainly based on “ring” shape,another was the one mainly based on “stylo” shape. The former one had typical crRNA after cleaving,while the function of the latter needs to be further studied. This paper laid the foundation for exploring the molecular mechanism of L. bulgaricus against phage and has great significance in screening anti-phage fermentative strains in the dairy industry via analyzing the CRISPR sequence of L. bulgaricus.

Key words: Lactobacillus bulgaricus, homology, CRISPR, spacer sequence, secondary structure

程娜,蔡针华,吕加平,贾震虎,逄晓阳. 保加利亚乳杆菌CRISPR位点的序列分析[J]. 生物技术通报, 2018, 34(5): 87-93.

CHENG Na, CAI Zhen-hua, LÜ Jia-ping, JIA Zhen-hu, PANG Xiao-yang. Sequence Analysis on the CRISPR Sites of Lactobacillus bulgaricus[J]. Biotechnology Bulletin, 2018, 34(5): 87-93.

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