猪源附红细胞体rnpB基因的克隆与序列比较  

CLONING AND SEQUENCE ANALYSIS OF RNPB GENE OF MYCOPLASMA SPP. ORIGIN OF SWINE

作　　者：王向佩[1,2] 周鹏[2] 米荣升[2] 沈莉萍[3] 黄燕[2] 王晓娟[1,2] 杨晓娇[2] 石凯[2] 刘宇轩[2] 雷晓思 陈兆国[2] 赵权[1] 

机构地区：[1]吉林农业大学动物科学技术学院,长春130118 [2]中国农业科学院上海兽医研究所农业部动物寄生虫学重点开放实验室中国农业科学院动物源性食品安全研究中心,上海200241 [3]上海市动物疫病预防控制中心,上海201103

出　　处：《中国动物传染病学报》2014年第3期26-33,共8页Chinese Journal of Animal Infectious Diseases

基　　金：公益性行业(农业)科研专项项目(200903036);中央级公益性科研院所基本科研业务费专项资金项目(2014JB04)

摘　　要：为了解猪源附红细胞体（Mycoplasma spp.）RNA酶P RNA（RNase P RNA,rnpB）基因序列变异状况,将实验室保存的经16S rRNA基因序列分析鉴定的53份猪源附红细胞体阳性DNA样品用于rnpB基因的扩增,扩增产物克隆至pMD18-T载体并进行序列测定。将获得的序列与GenBank登录的猪附红细胞体（Mycoplasma suis）德国分离株rnpB基因序列（登录号：EF523602）进行分析和比对,利用MEGA5.0软件构建系统发育树,并与16S rRNA判定结果进行比较。结果共得到42条猪附红细胞体rnpB基因和11条小附红细胞体（M.parvum）rnpB基因序列。猪附红细胞体上海分离株rnpB基因与GenBank登录的猪附红细胞体德国分离株rnpB基因之间的核苷酸序列相似性为98.1%～100.0%;小附红细胞体上海分离株rnpB基因与猪附红细胞体德国分离株rnpB基因序列的相似性为91.0%,与猪附红细胞体上海分离株rnpB基因序列相似性为90.0%～91.0%。系统进化分析显示猪附红细胞体rnpB基因与小附红细胞体rnpB基因分布在不同聚簇上,与16S rRNA基因序列鉴定结果一致。The present study was designed to illustrate variation of RNase P RNA （rnpB） gene of Mycoplasma spp. origin of swine. Partial rnpB gene of Mycoplasmasuis and M.parvum was amplified in PCR from 53 genomic DNAs extracted from pig blood samples, which had been identified as Mycoplasma spp. positive by 16S rRNA gene sequence analysis. The amplified products were cloned into pMD18-T vector for sequencing. The obtained sequences were compared with the sequence of rnpB gene of M. suis German isolate in GenBank （Accession number： EF523602）. The phylogenetic tree was constructed based on nucleotide sequence of rnpB gene of different species using MEGA 5.0 software and the results of Mycoplasma species identification were compared with those of 16S rRNA assessment. Total 42 rnpB genes of different M. suis isolates and 11 rnpB genes of different M. parvum isolates were cloned and sequenced. Homology analysis showed that the nucleotide sequence identities were 98.1%-100.0% between M. suis rnpB gene of Shanghai isolates and German isolate, 91.0%between rnpB gene of M. parvum Shanghai isolates and German isolate and 90.0%-91.0%between M.parvum and M.suis Shanghai isolates. The rnpB genes of M.suis and M.parvum distributed in different clusters of phylogenetic tree, which was consistent with 16S rRNA gene assessment.

关 键 词：猪附红细胞体 小附红细胞体 rnpB 克隆 序列比较 

分 类 号：S852.64[农业科学—基础兽医学]