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我国耳萝卜螺ITS及5.8S rDNA的克隆及序列分析

ISSN号：1673-4696
期刊名称：《中国兽医科学》
时间：0
分类：S852.735[农业科学—基础兽医学;农业科学—兽医学;农业科学—畜牧兽医]
作者机构：[1]华南农业大学兽医学院,广东广州510642, [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046, [3]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319
相关基金：公益性行业（农业）科研专项经费项目（201303037）; 甘肃省创新研究群体计划项目（1210RJIA006）

作者：王瑞爱[1,2], 刘国华[2], 王春仁[3], 张念章[2], 翁亚彪[1]

关键词：耳萝卜螺, 内转录间隔区, 核糖体DNA, 序列分析, 种系发育关系, Radix auricularia, ITS, ribosomal DNA, sequence analysis, phylogenetic relationship

中文摘要：

以采自我国黑龙江省大庆市的耳萝卜螺样品为研究对象,利用核糖体DNA（rDNA）内转录间隔区序列（ITS）的遗传标记特点,用保守引物BD1和BD2对rDNA内转录间隔区（ITS）及5.8S序列进行PCR扩增,扩增纯化后克隆至pMD18-T载体,重组质粒通过菌液PCR鉴定后,对阳性菌落进行测序和分析。结果显示,所获得的23个耳萝卜螺样品的ITS及5.8SrDNA序列总长度为1 124～1 130bp,存在一定差异,包含ITS-1（548～551bp）、5.8S（175bp）、ITS-2（399～404bp）序列。23个黑龙江省耳萝卜螺之间ITS-1序列的相似性在98.3%以上,ITS-2序列相似性在98.2%以上;而与GenBank中其他椎实螺的ITS-1序列的相似度低于75.6%,与ITS-2序列的相似度低于83.1%。由于耳萝卜螺ITS序列种内相对保守,种间差异较大,故可作为椎实螺种间遗传变异研究的标记。本研究结果为耳萝卜螺的分类鉴定以及进一步的分子流行病学研究奠定了基础。

英文摘要：

The objectives of the present study were to examine sequence variation in the internal transcribed spacers of ribosomal DNA（ITS rDNA） of Radix auricularia from Heilongjiang province,China.The ITS sequences were amplified from 23 R.auricularia samples and the amplicons were cloned into pMD18-T vector. The inserts were sequenced,and the lengths of ITS rDNA among 23 R.auricularia isolates was different（1124-1130 bp）.Sequence analysis revealed that the ITS-1,5.8S and ITS-2 rDNA of these samples were 548-551 bp,175 bp and 399-404 bp in size,respectively.Sequence similarities among the 23 R.auricularia samples were higher than 98.3% for ITS-1,and higher than 98.2% for ITS-2,while inter-specific difference among lymnaeid snails was considerable.The results of the present study indicate that ITS rDNA sequences can provide useful genetic markers for molecular identification of R.auricularia,and implications for further molecular epidemiological studies of R.auricularia.

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