霍乱弧菌山梨醇发酵相关基因序列特征分析

闫笑梅; 邹清华; 曾浔; 张建中

Abstract

闫笑梅,邹清华,曾浔,张建中.霍乱弧菌山梨醇发酵相关基因序列特征分析[J].Chinese journal of Epidemiology,2005,26(6):444-447

霍乱弧菌山梨醇发酵相关基因序列特征分析

Sequence analysis on sorbitol fermentation related genes in Vibrio cholerae

Received:January 17, 2005

DOI：

KeyWord: 霍乱弧菌山梨醇发酵基因测序

English Key Word: Vibrio cholerae Sorbitol fermention Gene sequencing

FundProject:科技部重大社会公益资助项目(2002DIA50036)

Author Name	Affiliation	E-mail
YAN Xiao-mei	National Institute of Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
ZOU Qing-hua	National Institute of Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
ZENG Xun	National Institute of Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
ZHANG Jian-zhong	National Institute of Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China	zhangjianzhong@icdc.cn

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Abstract:

目的比较霍乱弧菌流行株与非流行株山梨醇发酵相关基因的差异,为采用分子生物学方法快速区分两类菌株提供理论依据。方法采用基因测序的方法,分别对42株霍乱弧菌(O1群埃尔托型33株、O139群9株)流行株和非流行株的糖发酵激活蛋白、外周质麦芽糖结合蛋白、外周质磷酸盐结合蛋白和外周质氨基酸结合蛋白编码基因进行序列比较。结果糖发酵激活蛋白编码基因在霍乱弧菌流行株与非流行株共发现三个位置有单个碱基的差异,即第106、150和378位核苷酸(在流行株分别为A、A和T,而在非流行株分别为G、G和C)。第106位碱基的差异导致了氨基酸的不同(编码蛋白的第36个氨基酸在流行株和非流行株分别为苏氨酸和丙氨酸)。外周质麦芽糖结合蛋白和外周质磷酸盐结合蛋白编码基因各发现两个碱基的规律变化,外周质氨基酸结合蛋白编码基因未发现一致性碱基改变。结论在这些基因中存在着多个单核苷酸多态性,可能会成为快速区分两类菌株的重要依据。糖发酵激活蛋白第36位氨基酸的改变,可能会引起该蛋白活性的改变。

English Abstract:

Objective To I nvestig ate the differ ences of sorbitol fermentatio n related genes and optimize molecular analysis method for distinguishing an epidemic with no nepidemic strains of Vibrio cholerae. Methods Sequence analysis on four genes of sugar fermentation stimulation pr otein, periplasmic maltose binding protein, per iplasmic phosphate binding protein and periplasmic amino acid binding pr otein.Results In this study, the following data was noticed: for O1 serogroup El Tor bio type V. cholerae, twenty four epidemic and eight nonepidemic str ains were chosen; For O139 serogroup V. choler ae, five epidemic and four nonepidemic strains were chosen. With those g enes of sugar fermentation stimulation protein, there were thr ee point mutations. The 106th, 150th, 378th oligonucleot ide in epidemic str ains were A, A and T, comparing to the nonepidemic strains which w ere G, G and C. When comparing the protein sequences, epidemic str ains had a Threonine at 36th amino acid, w hereas nonepidemic strains had anAlanine. The r esults in O139 serogroup were consistent with those in O1 serog roup El Tor biotype strains.Another two point mutat ions were found in the genes of per iplasmic maltose binding protein. The 999th,1003th oligonucleotides in epidemic str ains were A and C, while in nonepidemic which were G and T. For the gene of periplasmic amino acid binding protein, two point mutatio ns w ere not iced. The 504th and 690th oligonucleotides in epidemic strains w ere T and C, but were C and T in nonepidemic. How ever, no aminoacid differences w ere found in periplasmic maltose binding protein and periplasmic amino acid binding protein. For per iplasmic amino acid binding protein g ene, there w as no difference on oligonucleotide between epidemic and nonepidemic strains. Conclusion Results suggested that SNPs in these genes might serve as a useful tool to distinguish the epidemic strains from nonepidemic strains. The 36th amino acid mutation of sugar fermentation stimulation protein in epidemic and nonepidemic strains mig ht change the activity of the pro tein w hich might be associated with sorbitol fermentation.

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