重庆地区牛附红细胞体16S rRNA基因部分序列的测定和分析

SEQUENCING AND PHYLOGENETIC ANALYSIS OF 16S RRNA GENE OF EPERYTHROZOON WENYONII IN CHONGQING

目的对牛附红细胞体的16SrRNA基因序列进行测定和系统进化分析。方法无菌采取感染附红细胞体的黄牛、奶牛、水牛血液,提取附红细胞体的基因组DNA,根据GenBank公布的奶牛附红细胞体16SrRNA序列设计的特异性引物进行PCR扩增,并对PCR产物进行序列测定和系统进化分析。结果PCR扩增出的DNA片段均为415bp左右。序列测定和系统进化分析显示,三者间的相似度分别为(黄牛:水牛=99.52%;黄牛:奶牛=99.28%;奶牛:水牛=99.76%)。与GenBank上公布的Mycoplasma wenyonii(武汉株)序列比较存在有4个突变位点,相似度均>98%。结论表明本次在重庆地区从牛体分离的附红细胞体为温氏附红细胞体。

The Eperythrozoon genomic DNA was extracted from blood samples of cattle,cow and buffalo. A pair of primers were designed based on the 16S rRNA sequence of Eperythrozoon wenyonii, which defined a fragment of about 415bp amplification product. The PCR products sequencing and phylogenetic analysis showed that the sequence from cattle, cow and buffalo were all highly similar to that of Mycoplasrna wenyonii in Wuhan with a homology of more than 98%, and the similarity between cattle and buffalo, cattle and cow, cow and buffalo was 99.52%, 99.28% and 99.76% respectively. The result indicated that the Eperythrozoon from the cattle, cow and buffalo in Chongqing district were E. wenyonii .