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基于胡子鲶(塘鲺)低深度基因组序列开发基因SSR标记并进行特征分析

Development and characterization of genic SSR markers from low depth genome sequence of Clarias batrachus (magur).

作者信息

Srivastava Shreya, Kushwaha Basdeo, Prakash Jyoti, Kumar Ravindra, Nagpure N S, Agarwal Suyash, Pandey Manmohan, Das P, Joshi C G, Jena J K

机构信息

Molecular Biology and Biotechnology Division, ICAR-National Bureau of Fish Genetic Resources, Lucknow 226 002,

出版信息

J Genet. 2016 Sep;95(3):603-9. doi: 10.1007/s12041-016-0672-8.

Abstract

Indian magur (Clarias batrachus) is an important freshwater catfish, which is listed as endangered under A3cde+4acde ver. 3.1 categories by the IUCN (2015) due to decreasing population trend. Microsatellites or short sequence repeats (SSRs) tagged to genes have been utilized as gene marker. In the present study, 31,814 SSRs of C. batrachus (magur) were identified using microsatellite identification tool programme from the next generation sequencing data generated on Roche 454 and Ion Torrent platforms. A bioinformatics pipeline, with stringent criteria resulted in selection of 1672 microsatellite loci falling in the genic region. Initially, a total of 30 loci were selected for primer development; and of these 14 were successfully amplified and five were found to be polymorphic in 30 individuals of C. batrachus (magur). The observed as well as expected heterozygosity ranged from 0.038 to 0.526 and 0.434 to 0.784, respectively, and the number of observed alleles ranged from three to five. The study reported the application of next generation sequencing technologies for rapid development of microsatellite loci in Indian catfish species, C. batrachus (magur).

摘要

印度胡子鲶(Clarias batrachus)是一种重要的淡水鲶鱼,由于其种群数量呈下降趋势,被国际自然保护联盟(2015年)列为A3cde + 4acde ver. 3.1类别下的濒危物种。与基因相关的微卫星或短序列重复序列(SSRs)已被用作基因标记。在本研究中,利用微卫星识别工具程序,从罗氏454和离子激流平台生成的下一代测序数据中,鉴定出31,814个印度胡子鲶(magur)的微卫星。一个具有严格标准的生物信息学流程,最终筛选出1672个位于基因区域的微卫星位点。最初,总共选择了30个位点用于引物开发;其中14个成功扩增,并且发现其中5个在30条印度胡子鲶(magur)个体中具有多态性。观察到的杂合度和预期杂合度分别在0.038至0.526和0.434至0.784之间,观察到的等位基因数量在3至5个之间。该研究报道了利用下一代测序技术在印度鲶鱼物种印度胡子鲶(magur)中快速开发微卫星位点的应用。

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