【摘要】 目的通过分析18SrRNA基因序列同源性,对河南猪株旋毛虫进行分子鉴定及分类。方法收集河南猪株旋毛虫成虫,提取总RNA,反转录合成cDNA,经特异引物扩增获得18SrRNA基因片段。将此目的基因与pMD18-T载体连接,转化大肠埃希菌感受态细胞,阳性克隆经PCR及酶切鉴定后进行序列测定及分析,构建系统发育树。结果构建的重组质粒酶切片段大小分别为2 700和1 800bp,与预期值相符。根据18SrRNA碱基序列构建系统发生树,河南猪株旋毛虫与虫株Trichinella nativa(AY487254.1)的亲缘关系较近,同源性为99.1%。结论河南猪株旋毛虫归属于T2。更多还原

【Abstract】 Objective To identify and classify Trichinella from swine in Henan Province at the molecular level by sequence homology analysis of the 18S rRNA gene.Methods Total RNA was extracted from adult Trichinella collected from swine in Henan.cDNA was obtained by reverse transcription.The 18S rRNA gene was amplified with a specific primer.The fragments of PCR products were ligated to pMD18-T.This was then transformed into E.coli competent cells.After identification by PCR and restrictive endonuclease digestion,the positive clone was sequenced and analyzed and then a phylogenetic tree was constructed.Results The fragments of the constructed recombinant plasmid were about 2 700 bp and 1 800 bp,which were consistent with expected values.In the phylogenetic tree based on the base sequence of the 18S rRNA gene,Trichinella from swine in Henan Province was the closest relative to T.nativa(AY487254.1) with sequence similarity of more than 99.1%.Conclusion Trichinella from swine in Henan Province was Trichinella nativa(T2).更多还原