一株精氨酸双水解酶阴性河弧菌的分子遗传学分析

发布时间：2018-01-07 17:18

本文关键词：一株精氨酸双水解酶阴性河弧菌的分子遗传学分析　出处：《疾病监测》2014年03期 　论文类型：期刊论文

【摘要】：目的赖氨酸、鸟氨酸脱羧酶阴性、精氨酸双水解酶阳性和阿拉伯糖发酵实验是区分河弧菌和其他非凝集弧菌的特征性化学指标。本研究对发现的1株精氨酸双水解酶阴性的河弧菌Ma-2531进行了分子遗传学分析。方法利用多对特异性引物对精氨酸双水解酶代谢通路的arc操纵子区基因簇进行普通聚合酶链反应(PCR)扩增,并对产物进行测序分析,同时比较Ma-2531和精氨酸双水解酶阳性菌株EF85003的生长曲线和培养液pH值的变化。结果针对arcBCA基因簇序列的3对引物arc-F/arc-R、arc-F/arc-rev和arc-ck-up/arc-R均为阴性扩增,而其上下游序列特异性的引物对arc1-up/arc1-dn和arc72695-up/arc75219-dn分别扩增出预期大小的目的条带。arc1-dnRev/arc72695-upRev引物对的扩增产物序列分析显示该扩增片段包含transposase IS4序列,进一步的对比分析表明包括精氨酸双水解酶系统arcBCAD基因簇及上游的天冬氨酸氨甲酰转移酶调节亚基和催化亚基编码基因在内的约8.6 kb的大片段发生了缺失。生长曲线和pH值测定显示菌株Ma-2531和EF85003没有明显差异。结论菌株Ma-2531中插入序列的转座导致了该菌株精氨酸双水解酶这一鉴定生化指标阴性的结果,但这种变化应只是随机和罕见的,使河弧菌生化代谢表现出多样性。
[Abstract]:Objective Lysine and ornithine decarboxylase were negative. Arginine dihydrolase positive and arabinose fermentation were the characteristic chemical indexes to distinguish Vibrio river from other non agglutination vibrio. In this study, a strain of Ma-2531 with arginine dihydrolase negative was studied. Methods the arc operon gene cluster of arginine dihydrolase metabolic pathway was analyzed by polymerase chain reaction (PCR) with multiple pairs of specific primers. PCR amplification. The products were sequenced and analyzed. At the same time, the growth curve of Ma-2531 and arginine double hydrolase positive strain EF85003 and the change of pH value of culture medium were compared. Results three pairs of primers arc-. Frarc-R. Arc-F/arc-rev and arc-ck-up/arc-R were both negative amplification. The desired size of the target band. Ar was amplified from arc1-up/arc1-dn and arc72695-up/arc75219-dn by the upstream and downstream sequence specific primers, respectively. Sequence analysis of c1-dnRevarc72695-upRev primer pair showed that the amplified fragment contained transposase. IS4 sequence. Further comparative analysis showed that about 8.6 genes, including the arcBCAD gene cluster of arginine dihydrolase system and the upstream aspartic carbamyl transferase regulatory and catalytic subunit coding genes, were included in this study. The growth curve and pH test showed that there was no significant difference between Ma-2531 and EF85003. Conclusion the transposition of insertion sequence in Ma-2531 results in the translocation of the insertion sequence in strain Ma-2531. The identification of arginine dihydrolase was negative. However, these changes should be random and rare, resulting in diversity of biochemical metabolism of Vibrio river.
【作者单位】：中国疾病预防控制中心传染病预防控制所;
【基金】：国家自然科学基金(No.81071410) “十二五”国家科技重大专项课题(No.2013ZX10004-101002)~~
【分类号】：R378.2
【正文快照】： 河弧菌(Vibrio fluvialis)是一种新发的食源性致病菌,1975年首次分离于巴林1名腹泻患者粪便中,也曾被称为F群弧菌(Group F vibrios)和EF-6弧菌(EF-6 vibrios)[1-3]。河弧菌自然分布于温暖含盐的水体中,可通过污染海产品、淡水及海水水体得以传播。在人类主要引起急性胃肠炎,

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