用PCR-RFLP技术鉴别鲁道夫对盲囊线虫姊妹种的研究  

Genetic identification of Contracaecum rudolphii complex by PCR-RFLP

作　　者：何芳[1] 屈仁建 吴绍强[1] 宋慧群[1] 林瑞庆[1] 朱兴全[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]山西恒丰强动物药业有限公司广州分公司,广东广州510650

出　　处：《华南农业大学学报》2005年第2期115-117,共3页Journal of South China Agricultural University

基　　金：国家杰出青年科学基金项目 (30 2 2 5 0 33)

摘　　要：用保守引物扩增鲁道夫对盲囊线虫Contracaecumrudolphii姊妹种和C .septentrionalerDNA第一内转录间隔区(ITS 1)片段并纯化,根据鲁道夫对盲囊线虫A、鲁道夫对盲囊线虫B和C .septentrionalerDNAITS 1序列,选用限制性内切酶MspΙ和NsiI酶切,酶切产物用琼脂糖凝胶电泳分析.结果经MspΙ酶切后,鲁道夫对盲囊线虫姊妹种与C .septentrionale表现出不同的条带;经NsiΙ酶切后,鲁道夫对盲囊线虫B和C .septentrionale结果一致,与鲁道夫对盲囊线虫A不同.用MspΙ可以鉴定出C .septentrionale ,用NsiΙ可以鉴定出鲁道夫对盲囊线虫A ,2个限制性内切酶合用可以将鲁道夫对盲囊线虫A、鲁道夫对盲囊线虫B以及C .septentrionale分别鉴定出来.根据鲁道夫对盲囊线虫A、鲁道夫对盲囊线虫B以及C .septentrionalerDNAITS 1基因序列建立的鲁道夫对盲囊线虫姊妹种PCR RFLP鉴别技术能够对鲁道夫对盲囊线虫姊妹种进行准确。Using a pair of conserved primers, the first internal transcribed spacer (ITS-1) of ribosomal DNA (rDNA) of Contracaecum rudolphii complex and C. septentrionale were amplified by PCR. The purified amplicons were digested by Msp Ι and Nsi Ι selected according to the ITS-1 sequences of C. rudolphii A, C. rudolphii B and C. septentrionale, followed by agarose gel electrophoresis analyses. The banding profiles after digestion with Msp Ι were different between the C. rudolphii complex and C. septentrionale, and the profiles of C. rudolphii B digested by Nsi I was identical to that of C. septentrionale, but different from that of C. rudolphii A. In combination of using Msp Ι and Nsi I, C. rudolphii A, C. rudolphii B and C. septentrionale could be unequivocally differentiated. The PCR-RFLP approach was simple, specific and provided a useful molecular tool for the identification of the C. rudolphii A, C. rudolphii B and C. septentrionale.

关 键 词：PCR-RFLP 鲁道夫对盲囊线虫 姊妹种 RDNA 第一内转录间隔区 鉴别技术 

分 类 号：S852.731[农业科学—基础兽医学]