中国动物传染病学报
中國動物傳染病學報
중국동물전염병학보
CHINESE JOURNAL OF VETERINARY PARASITOLOGY
2014年
3期
26-33
,共8页
王向佩%周鹏%米荣升%沈莉萍%黄燕%王晓娟%杨晓娇%石凯%刘宇轩%雷晓思%陈兆国%赵权
王嚮珮%週鵬%米榮升%瀋莉萍%黃燕%王曉娟%楊曉嬌%石凱%劉宇軒%雷曉思%陳兆國%趙權
왕향패%주붕%미영승%침리평%황연%왕효연%양효교%석개%류우헌%뢰효사%진조국%조권
猪附红细胞体%小附红细胞体%rnpB%克隆%序列比较
豬附紅細胞體%小附紅細胞體%rnpB%剋隆%序列比較
저부홍세포체%소부홍세포체%rnpB%극륭%서렬비교
Mycoplasma suis%M.parvum%rnpB gene%cloning%sequence alignment
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。
為瞭解豬源附紅細胞體（Mycoplasma spp.）RNA酶P RNA（RNase P RNA，rnpB）基因序列變異狀況，將實驗室保存的經16S rRNA基因序列分析鑒定的53份豬源附紅細胞體暘性DNA樣品用于rnpB基因的擴增，擴增產物剋隆至pMD18-T載體併進行序列測定。將穫得的序列與GenBank登錄的豬附紅細胞體（Mycoplasma suis）德國分離株rnpB基因序列（登錄號：EF523602）進行分析和比對，利用MEGA5.0軟件構建繫統髮育樹，併與16S rRNA判定結果進行比較。結果共得到42條豬附紅細胞體rnpB基因和11條小附紅細胞體（M.parvum）rnpB基因序列。豬附紅細胞體上海分離株rnpB基因與GenBank登錄的豬附紅細胞體德國分離株rnpB基因之間的覈苷痠序列相似性為98.1%~100.0%；小附紅細胞體上海分離株rnpB基因與豬附紅細胞體德國分離株rnpB基因序列的相似性為91.0%，與豬附紅細胞體上海分離株rnpB基因序列相似性為90.0%~91.0%。繫統進化分析顯示豬附紅細胞體rnpB基因與小附紅細胞體rnpB基因分佈在不同聚簇上，與16S rRNA基因序列鑒定結果一緻。
위료해저원부홍세포체（Mycoplasma spp.）RNA매P RNA（RNase P RNA，rnpB）기인서렬변이상황，장실험실보존적경16S rRNA기인서렬분석감정적53빈저원부홍세포체양성DNA양품용우rnpB기인적확증，확증산물극륭지pMD18-T재체병진행서렬측정。장획득적서렬여GenBank등록적저부홍세포체（Mycoplasma suis）덕국분리주rnpB기인서렬（등록호：EF523602）진행분석화비대，이용MEGA5.0연건구건계통발육수，병여16S rRNA판정결과진행비교。결과공득도42조저부홍세포체rnpB기인화11조소부홍세포체（M.parvum）rnpB기인서렬。저부홍세포체상해분리주rnpB기인여GenBank등록적저부홍세포체덕국분리주rnpB기인지간적핵감산서렬상사성위98.1%~100.0%；소부홍세포체상해분리주rnpB기인여저부홍세포체덕국분리주rnpB기인서렬적상사성위91.0%，여저부홍세포체상해분리주rnpB기인서렬상사성위90.0%~91.0%。계통진화분석현시저부홍세포체rnpB기인여소부홍세포체rnpB기인분포재불동취족상，여16S rRNA기인서렬감정결과일치。
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.
