单增李斯特菌新疆分离株lmo2192基因克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.11.005

摘要/Abstract

摘要：

试验旨在对单增李斯特菌新疆绵羊脑炎临床分离株LM90SB2的lmo2192基因进行克隆及生物信息学分析。根据GenBank中lmo2192基因序列（登录号：CAD00270）设计特异性引物，利用PCR方法对lmo2192基因进行扩增，回收目的基因，连接到pMD19-T载体上进行克隆，筛选阳性菌进行测序，测序后对lmo2192基因核苷酸序列进行分析，预测其编码蛋白质的二级结构、三级结构，对其进行同源性比对及遗传变异分析。结果显示，新疆分离株LM90SB2的lmo2192基因序列全长为1 277 bp，包含969 bp开放阅读框，共编码322个氨基酸；LM90SB2株lmo2192基因核苷酸序列与CⅡMS-PH-1同源性为100.0%，与81-0861、10-0809、81-0592、81-0558、NTSN、F2365和WSLC1033的同源性为99.8%~99.9%，与L2074和NH1同源性分别为97.0%和96.9%。推导的氨基酸序列同源性为91.6%~100.0%。系统进化树显示，LM90SB2菌株lmo2192基因与4b血清型菌株亲缘关系较近，聚为同一分支。蛋白质二级结构预测表明，LM90SB2 lmo2192蛋白为亲水性蛋白，无信号肽，不形成跨膜结构。蛋白结构域预测，lmo2192蛋白为ATP酶组分。本试验成功克隆LM90SB2分离株lmo2192基因，为进一步研究其基因功能提供理论依据。

关键词: 单增李斯特菌; lmo2192基因; 克隆; 生物信息学分析

Abstract:

The study was aimed to clone and bioinformatics analysis of lmo2192 gene of Listeria monocytogenes LM90SB2 isolated from diseased sheep in Xinjiang.Specific primers were designed according to the lmo2192 gene sequence in GenBank (accession No.:CAD00270).The PCR method was used to amplify lmo2192 gene.The PCR products were purified and cloned into pMD19-T vector,and then the positive colonies were screened and sequenced,and the nucleotide sequence of lom2192 gene was sequenced and analyzed.The secondary and tertiary structures of the encoded proteins were predicted,and homology and genetic variation analysis were performed.The results showed that LM90SB2 lmo2192 gene was 1 277 bp in length,containing a 969 bp open reading frame (ORF) that encoded 322 amino acids.The nucleotide sequence homology of lmo2192 gene of LM90SB2 strain was 100.0% with CⅡMS-PH-1;99.8% to 99.9% with 81-0861,10-0809,81-0592,81-0558,NTSN,F2365 and WSLC1033;And 97.0% and 96.9% with L2074 and NH1,respectively.The putative amino acid sequence homology was 91.6% to 100.0% with above strains.The phylogenetic tree showed that the lmo2192 gene of LM90SB2 strain was closely related to the serotype 4b strain and clustered into the same branch.The protein secondary structure prediction result showed that LM90SB2 lmo2192 protein was a hydrophilic protein without signal peptide and transmembrane structure.The protein domain predicted result showed that the lmo2192 protein was an ATPase component.The lmo2192 gene of LM90SB2 strain was successfully cloned,it provided theoretical basis for further study of the lmo2192 gene function of LM90SB2.

Key words: Listeria monocytogenes; lmo2192 gene; cloning; bioinformatics analysis

中图分类号:

S852.61

李红欢, 钱凌霄, 杜冬冬, 张奇文, 李庆辉, 马勋. 单增李斯特菌新疆分离株lmo2192基因克隆及生物信息学分析[J]. 《中国畜牧兽医》, 2018, 45(11): 3011-3017.

LI Honghuan, QIAN Lingxiao, DU Dongdong, ZHANG Qiwen, LI Qinghui, MA Xun. Cloning and Bioinformatics Analysis of lmo2192 Gene of Listeria monocytogenes from Xinjiang[J]. , 2018, 45(11): 3011-3017.

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