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我国犬钩虫ITS及5.8S rDNA的克隆与序列分析

Cloning and sequence analysis of the ITS and 5.8S rDNA of Ancylostoma caninum in China

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【作者】 宋慧群李宾林瑞庆朱兴全

【Author】 SONG Hui-qun,LI Bin,LIN Rui-qing,ZHU Xing-quan(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)

【机构】 华南农业大学兽医学院华南农业大学兽医学院 广东广州510642广东广州510642

【摘要】 以从我国广东湛江犬小肠中采集的2条犬钩虫作为研究对象,用保守引物NC5及NC2扩增犬钩虫rDNA的ITS-1、5.8S及ITS-2片段,将PCR扩增出的片段纯化后克隆至pGEM-T Easy载体,重组质粒通过菌落PCR和酶切鉴定后,对阳性菌落进行序列测定并进行序列分析。结果显示,来自湛江的2条犬钩虫ITS及5.8S rDNA序列总长均为738 bp,其中ITS-1序列长为364 bp,5.8S序列长为153 bp,ITS-2序列长为221 bp;2个不同样品之间ITS序列存在多态性,ITS-1序列存在5个碱基的差异,ITS-2序列存在1个碱基的差异,5.8S rDNA序列无差异。研究结果为犬钩虫进一步的分类、鉴定和遗传变异研究奠定了基础。

【Abstract】 The internal transcribed spacer(ITS) and 5.8S rDNA of Ancylostoma caninum isolated in Guangdong Province of China were amplified by PCR using a pair of conserved primers and the amplicons were cloned into pGEM-T Easy vector,respectively.The inserts were successfully sequenced,and the ITS of the two A.caninum samples was 738 bp in length.Sequence analysis revealed that the ITS-1,5.8S and ITS-2 rDNA of both A.caninum samples were 364 bp,153 bp and 221 bp in length,respectively,and polymorphism was found in the ITS-1 and ITS-2 rDNA sequences between the two A.caninum samples.The results provided a foundation for further studies of molecular identification and molecular genetics of A.caninum.

【基金】 国家杰出青年科学基金项目(30225033)
  • 【文献出处】 中国兽医科学 ,Veterinary Science in China , 编辑部邮箱 ,2007年09期
  • 【分类号】S852.7
  • 【被引频次】12
  • 【下载频次】320
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