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IRAP:一种基于反转录转座子的高效电泳技术,用于研究日本血吸虫地理株间的遗传变异。

IRAP: An efficient retrotransposon-based electrophoretic technique for studying genetic variability among geographical isolates of Schistosoma japonicum.

机构信息

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu Province, P R China.

出版信息

Electrophoresis. 2011 Jun;32(12):1473-9. doi: 10.1002/elps.201100044. Epub 2011 May 30.

Abstract

In the present study, a inter-retrotransposon-amplified polymorphism (IRAP) technique, based on retrotransposons, was used to examine genetic variability among Schistosoma japonicum isolates from different provinces in mainland China. Of the 15 primers screened, 5 produced highly reproducible IRAP patterns. Using these primers, 54 discernible DNA fragments were generated with 40 (74.07%) being polymorphic, indicating considerable genetic variation among the examined S. japonicum isolates. The primer LTR-11 was found to be able to differentiate male and female parasites, producing one constant specific band for female S. japonicum isolates. The percentages of polymorphic bands (PPB) among all parasites, among isolates from mountainous provinces and among those from the lake/marshland areas were 74.07, 48.15, and 66.67%, respectively. UPGMA analysis revealed that the IRAP profiles could group S. japonicum isolates in mainland China into two clades (mountainous and lake/marshland types), and samples from the same geographical origins clustered together. These results demonstrated that the IRAP technique is suitable for studying genetic diversity and population structures, and also provides an effective technique for studying sex differentiation of S. japonicum.

摘要

在本研究中,我们使用一种基于逆转座子的反转录转座子扩增多态性(IRAP)技术,研究了中国大陆不同省份间日本血吸虫分离株的遗传变异。在筛选出的 15 个引物中,有 5 个产生了高度可重复的 IRAP 图谱。利用这些引物,共产生了 54 个可分辨的 DNA 片段,其中 40 个(74.07%)具有多态性,表明所研究的日本血吸虫分离株存在相当大的遗传变异。引物 LTR-11 被发现能够区分雌雄寄生虫,对雌性日本血吸虫分离株产生一个恒定的特异带。所有寄生虫、山区分离株和湖区/沼泽地分离株的多态性带百分比(PPB)分别为 74.07%、48.15%和 66.67%。UPGMA 分析表明,IRAP 图谱可以将中国大陆的日本血吸虫分离株分为两个分支(山区和湖区/沼泽地类型),并且来自同一地理起源的样本聚集在一起。这些结果表明,IRAP 技术适合于研究遗传多样性和种群结构,也为研究日本血吸虫的性别分化提供了一种有效技术。

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