SpHtp1基因在不同种属水霉菌株中的分布
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国家“863”项目(2011AA10A216)、公益性行业(农业)科研专项(201203085)、上海市科技兴农重点攻关项目(沪农科攻字(2011)第4-8号)、国家自然科学基金项目(31172430)、国家水产种质资源平台运行服务项目和上海高校知识服务平台项目


Distribution of SpHtp1 gene among different Saprolegnia strains
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    摘要:

    为研究SpHtp1基因在不同种属水霉菌株中的分布,以寄生水霉ATCC200013TM的基因组DNA为模板,设计1对特异性引物,进行PCR扩增。将扩增片段回收,并克隆入T载体,转化大肠杆菌DH5α,挑取阳性克隆测序。将测序结果与GenBank中登录的SpHtp1基因序列进行比较,同源性为99%。以该方法调查从水霉病主要流行地区分离的17株水霉菌SpHtp1基因的分布情况,结果发现:在17株水霉菌株中,有8株水霉菌检测到SpHtp1基因,阳性率为47%,且所有含SpHtp1基因的菌株在分类地位上均隶属于寄生水霉(Saprolegnia parasitica)。 SpHtp1基因可能在寄生水霉中特异性存在,该基因有望用于寄生水霉的分离鉴定。

    Abstract:

    In order to understand the distribution of the SpHtp1 gene among different saprolegnia strains,using Saprolegnia parasitica (ATCC200013TM) genomic DNA as the template,a pair of gene-specific primers was designed,a PCR method was developed and the parameters for PCR amplification was optimized.The PCR product was extracted,linked into pMD18-T vector and then cloned into E.coli DH5α.The recombination plasmid was identified by PCR and sequenced to prove the amplification of the target gene.As a result,a 224 bp DNA fragment of the SpHtp1 gene can be amplified from the saprolegnia genome,which had an identity of 99% in sequence with that of other Saprolegnia strains.Additionally,17 samples from all over China were tested using this method and the SpHtp1 gene was present in 8 (47%) samples.The results indicated that the SpHtp1 gene is expressed specifically in S.parasitica,which can be used potentially for detection of Saprolegnia.

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叶鑫,赵依妮,曹海鹏,胡鲲,杨先乐. SpHtp1基因在不同种属水霉菌株中的分布[J].华中农业大学学报,2014,33(05):99-104

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  • 收稿日期:2013-12-10
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