nd-fish 技术在两栖类中的应用.pdfVIP

动 物 学 研 究 2013，Dec. 34(6): 610−616 CN 53-1040/Q ISSN 0254-5853 Zoological Research DOI:10.11813/j.issn.0254-5853.2013.6.0610 ND-FISH 技术在两栖类中的应用 1,2 1 1,* 常晓嫒 ，夏 云 ，曾晓茂 1. 中国科学院成都生物研究所，四川 成都 610041 2. 中国科学院大学，北京 100049 摘要：非变性荧光原位杂交（ND-FISH ）是近年来兴起的一种技术，一般使用带有荧光标记的简单序列重复（SSRs ）作为 探针，杂交中不需要进行探针和染色体的变性使得检测简单快捷。SSRs 探针可检测出染色体上SSR 富集的区域，可以帮助 辨识染色体，SSRs 分布的多态性还可以为遗传多态性的研究提供信息。（1）该研究首次将ND-FISH 运用于两栖动物中，得 到了5 个可运用于多态性研究的标记。（2 ）研究发现，有别于其他类群，当ND-FISH 应用于两栖动物时，染色体标本必须 进行变性。（3 ）该研究还对双色荧光原位杂交流程进行了较大的改进，在一个杂交体系中同时使用直接和间接标记探针，通 过一次实验获得双色信号，从而使步骤简化。 关键词：ND-FISH ；SSRs；标记筛选；双色FISH 中图分类号：Q959.5; Q7; Q343 文献标志码：A 文章编号：0254-5853-(2013)06-0610-07 Application of ND-FISH in amphibians 1,2 1 1,* Xiao-Ai CHANG , Yun XIA , Xiao-Mao ZENG 1. Chengdu Institute of Biology Chinese Academy of Science, Chengdu 610041, China 2. University of Chinese Academy of Science, Beijing 100049, China Abstract: The recently popularized non-denaturing fluorescence in situ hybridization (ND-FISH) is a new technique that is both quick and efficient, in part because denaturing of both of the probes and the chromosomes is unnecessary. Synthetic simple sequence repeats (SSRs) labeled with fluorescein are used as probes to detect SSR-enriched chromosome regions and provide markers to identify the chromosomes. To date this method has not been applied to amphibians, even though the polymorphism of the distribution of SSRs may help to advance genetic polymorphism research. This paper also improved the double-colour FISH method by simultaneously using probes labelled with fluorescein and probes labelled with DIG to get double-color signals. This study found 5 SSRs markers that may