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胭脂鱼微卫星富集文库的构建及其应用前景展望

Construction of Microsatellite Library Enriched and its Application Prospect in Chinese sucker(Myxocyprinus asiaticus)

投稿时间：2009-03-02

DOI：

中文关键词:胭脂鱼微卫星富集文库

英文关键词:Chinese sucker microsatellite enriched library

基金项目:

作者	单位
杨钟	水利部中国科学院水工程生态研究所，湖北武汉
史芳	水利部中国科学院水工程生态研究所，湖北武汉，中国科学院水生生物研究所，湖北武汉
阙延福	水利部中国科学院水工程生态研究所，湖北武汉
熊美华	水利部中国科学院水工程生态研究所，湖北武汉
徐念	水利部中国科学院水工程生态研究所，湖北武汉，中国科学院水生生物研究所，湖北武汉
朱滨	水利部中国科学院水工程生态研究所，湖北武汉

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中文摘要:

以湖北武汉和四川宜宾人工增殖放流的胭脂鱼子一代酒精浸泡鳍条样本为材料，提取完整基因组ＤＮＡ，选用人工合成的生物素标记（ＡＡＡＧ）７探针，采用ＦＩＡＳＣＯ（fast isolation by AFLP of sequence containing repeats）磁珠富集法构建胭脂鱼（Ｍｙｘｏｃｙｐｒｉｎｕｓａｓｉａｔｉｃｕｓ）微卫星富集文库。以ＭｓｅＩ－Ｎ引物组和设计合成的微卫星核心序列引物（ＡＡＡＧ）５，用双引物ＰＣＲ法筛选有微卫星的阳性克隆，从５４个阳性克隆中共获得２２个微卫星序列，其中完美型（ｐｅｒｆｅｃｔ）共１５个（占６８．２％），非完美型（ｉｍｐｅｒｆｅｃｔ）６（占２７．３％），混合型（ｃｏｍｐｏｕｎｄ）１个（占４５％）；（ＡＡＡＧ／ＴＴＴＣ）ｎ序列在胭脂鱼的基因组ＤＮＡ中含量非常丰富。根据微卫星侧翼序列最终设计并合成了１８对胭脂鱼微卫星引物，经其有效性检验后，可应用于胭脂鱼遗传多样性、种群遗传结构等进一步研究及人工增殖放流效果评估。

英文摘要:

This research gets the fins of Chinese sucker from Wuhan city of Hubei province and Yibin city of Sichuan province, which are belong to releasing generation one. We have extracted the whole genomic DNA from these tissues. The enriched microsatellite library of Chinese sucker were constructed by FIASCO （ Fast Isolation by AFLP of Sequences containing repeats） magnetic beads method, using the biotin-labeled probe （AAAG）7 by artificial synthesization. 22 microsatellites in 54 recombinant positive clones were obtained through PCR screening the li- brary with MseI-N primers and simple tandem repeats primer （AAAG）5- Among these microsatellites, there were 15 perfect ones （68.2%） ,6 imperfect ones （27.3%） and 1 compound ones （4.5%）. The results indicated that mi- crosatellite sequences characterized by （ AAAG/TTTC）, were abundant in genomic DNA of Chinese sucker. Among the 22 microsatellite sequences, 18 pairs of primers were designed to analyze genomic DNA of Chinese sucker according to unique microsatellite flanking sequences with the software Primer3, Premier primer 5.0 and Oligo 6. After testing their validity, these microsatellite primers can be used to further study the genetic diversity, population genetic structure and assess the artificial propagation releasing effect in Chinese sucker.

杨钟,史芳,阙延福,熊美华,徐念,朱滨.2009.胭脂鱼微卫星富集文库的构建及其应用前景展望[J].水生态学杂志,30(2):107-112.

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