短小乳杆菌内蒙古分离株S-层蛋白的普查、鉴定和基因分析

中国畜牧兽医

短小乳杆菌内蒙古分离株S-层蛋白的普查、鉴定和基因分析

兰丽^1,2, 魏建民², 王艳霞¹, 王敏¹, 满达¹, 格日勒图¹

1. 内蒙古农业大学乳品生物技术与工程教育部重点实验室,内蒙古呼和浩特 010018;
2. 内蒙古农业大学生命科学学院,内蒙古呼和浩特 010018

收稿日期:2012-03-12 出版日期:2012-08-20 发布日期:2012-08-24
通讯作者: 格日勒图 E-mail:geriletunjau@yahoo.com.cn
作者简介:兰丽(1985-),女,内蒙古人,硕士生,研究方向:蛋白质化学。
基金资助:
国家自然基金项目(30960279);内蒙古自然基金项目(2011BS0408)。

Detection,Identification and Genetic Analysis of S-Layer Protein of Lactobacillus brevis Isolated from Inner Mongolia

LAN Li^1,2, WEI Jian-min², WANG Yan-xia¹, WANG Min¹, MAN Da¹, Geriletu¹

1. Ministry of Education Key Laboratory of Dairy Biotechnology and Engineering, Inner Mongolia Agricultural University, Hohhot 010018,China;
2. College of Life Sciences,Inner Mongolia Agricultural University,Hohhot 010018,China

Received:2012-03-12 Online:2012-08-20 Published:2012-08-24

摘要/Abstract

摘要： S-层蛋白(S-layers protein)是位于某些细菌表面的高度有序的具有超微结构的一种外膜蛋白。本试验以蒙古族传统乳制品中分离的短小乳杆菌(Lactobacillus brevis)为材料,应用SDS-PAGE技术对若干乳杆菌进行S-层蛋白的普查,并应用PCR方法克隆出L.brevis的slp基因,对该slp基因进行测序及序列分析。试验结果表明,SDS-PAGE电泳结果显示L.brevis样品在44~55 ku之间出现目的条带,与文献报道的S-层蛋白大小范围一致;PCR方法正确扩增出大小约为1300 bp的slp基因, 经DNAStar软件遗传分析发现该基因与鸡乳酸杆菌株(GenBank登录号:AY597266.1)的亲缘关系和遗传距离最近,与其他种类乳酸菌遗传距离也非常相近;经Genetyx生物软件推导出的S-层蛋白大小理论值约为47 ku,具有显著的外膜蛋白的特性。本试验为后续遗传改造等研究工作奠定基础。

关键词: 短小乳杆菌; S-层蛋白; slp基因; 克隆与序列分析

Abstract: The surface layers protein (SLP) is a major composition of the bacterial outermost envelope with highly ordered ultrastructure. The Lactobacillus brevis which isolated from Mongolian traditional dairy products were used as materials in this study. The S-layer proteins from several Lactobacillus were detected by SDS-PAGE and slp gene was cloned by PCR and then analyzed the slp gene sequence of Lactobacillus breris. The results showed that the protein sample of Lactobacillus breris showed target band about 44 to 55 ku which had the same size range of SLP that had been reported,and slp gene about 1300 bp was amplified by PCR method. The DNAStar software found that the protein had recently kinship and genetic distance with L.gallinarum strain (GenBank:AY597266.1) and was similar to other Lactobacillus spp. The theoretical size of S-layer protein was about 47 ku released by the Genetyx software and had significant feature of outer membrane proteins. This layed foundation for genetic modification and other research work.

Key words: Lactobacillus brevis; S-layer protein; slp gene; cloning and sequence analysis

中图分类号:

Q781

兰丽, 魏建民, 王艳霞, 王敏, 满达, 格日勒图. 短小乳杆菌内蒙古分离株S-层蛋白的普查、鉴定和基因分析[J]. 中国畜牧兽医.

LAN Li, WEI Jian-min, WANG Yan-xia, WANG Min, MAN Da, Geriletu. Detection,Identification and Genetic Analysis of S-Layer Protein of Lactobacillus brevis Isolated from Inner Mongolia[J]. .

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