| 嗜热链球菌CRISPR-Cas系统的检测 |
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| 引用本文： | 李婉,梁宏彰,张丹青,王娜娜,唐雅茹,李柏良,霍贵成.嗜热链球菌CRISPR-Cas系统的检测[J].微生物学报,2016,56(4):680-688. |
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| 作者姓名： | 李婉 梁宏彰 张丹青 王娜娜 唐雅茹 李柏良 霍贵成 |
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| 作者单位： | 东北农业大学, 乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030,东北农业大学, 乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030,东北农业大学, 乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030,东北农业大学, 乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030,东北农业大学, 乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030,东北农业大学, 乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030,东北农业大学, 乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030 |
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| 基金项目： | 国家自然科学基金(31171717);国家"863计划"(2012AA022108) |
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| 摘 要： | 【目的】CRISPR-Cas系统为嗜热链球菌抵抗噬菌体等外源基因元件提供获得性免疫,分析NCBI中已公开发表全基因组序列的9株嗜热链球菌所含CRISPR-Cas系统的数目和类型,对实验室相应菌株的CRISPR-Cas系统进行检测。【方法】利用生物信息学方法对NCBI中9株已测序嗜热链球菌所含CRISPR-Cas系统进行分析,根据其Cas基因序列设计引物,对实验室嗜热链球菌菌株的Cas基因进行扩增、测序,分析实验室6株嗜热链球菌的CRISPR-Cas系统情况。【结果】9株标准菌株均含不同数目的CRISPR-Cas系统,其类型主要为Ⅱ-A型、Ⅲ-A型和Ⅰ-E型,各类型的标志Cas基因高度保守。6株供试菌中,S4仅含Cas9基因,其它5株均含有Cas9基因、Cas10基因和Cas9*基因,79和KLDS3.0207还含有Cas3基因。【结论】可根据标准菌株高度保守的Cas基因设计引物,预测未知嗜热链球菌所含CRISPRCas系统的数目和类型。S4仅含1个Ⅱ-A型CRISPR-Cas系统,其它5株均含有2个Ⅱ-A型CRISPR-Cas系统和1个Ⅲ-A型CRISPR-Cas系统,此外,79和KLDS3.0207均含有1个Ⅰ-E型CRISPR-Cas系统。
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| 关 键 词： | 嗜热链球菌 规律成簇的间隔回文重复(CRISPR) CRISPR-Cas系统 Cas 生物信息学分析 |
| 收稿时间： | 2015/7/20 0:00:00 |
| 修稿时间： | 9/9/2015 12:00:00 AM |
Detection of CRSPR-Cas system in Streptococcus thermophilus |
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| Wan Li,Hongzhang Liang,Danqing Zhang,Nana Wang,Yaru Tang,Bailiang Li and Guicheng Huo.Detection of CRSPR-Cas system in Streptococcus thermophilus[J].Acta Microbiologica Sinica,2016,56(4):680-688. |
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| Authors: | Wan Li Hongzhang Liang Danqing Zhang Nana Wang Yaru Tang Bailiang Li and Guicheng Huo |
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| Institution: | Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China,Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China,Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China,Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China,Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China,Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China and Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China |
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| Abstract: | Objective] We aimed to detect the CRSPR-Cas system of six Streptococcus thermophilus. Methods] Bioinformatics method was used to predict CRSPR-Cas system of nine S. thermophilus that published in National Center for Biotechnology Information. Four primers were designed according to the flanking sequences of standard strains and the CRISPR-Cas system of six S. thermophilus have been detected by PCR method. Results] S. thermophilus S4 had a Cas9 gene, others all had Cas9 gene, Cas10 gene and Cas9* gene. In addition, 79 and KLDS3.0207 still had Cas3 gene. Conclusion] Signature genes amplification of CRSPR-Cas system could predict the type of CRSPR-Cas system in unsequenced strains, these findings will help establish the foundation for the study of CRSPR-Cas system in lactic acid bacteria. |
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| Keywords: | Streptococcus thermophilus clustered regularly interspaced short palindromic repeats(CRISPR) CRSPR-Cas system Cas genes bioinformatics method |
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