[an error occurred while processing this directive]
en

Enrichment and screening of the microsatellite markers in Microtus fortis and its preliminary application in genetic diversity research

Expand
  • 1. Shanghai Laboratory Animal Research Center, Shanghai 201203, China 2. Shanghai Quality Monitoring Center for Laboratory Animal, Shanghai 201203, China 3. Shanghai SIPPR-BK LAB Animal ltd, Shanghai 201203, China

Received date: 2010-12-22

  Revised date: 2011-06-03

  Online published: 2011-09-25

Abstract

This study was to isolate microsatellite markers from Microtus fortis genome by magnetic beads enrichments. Through hybridization of biotin-labeled microsatellite oligonucleotide probes, which were captured by streptavidin-coated magnetic with the adaptor–ligated enzyme-digested genome fragments, single-stranded DNA fragments containing mi-crosatellites were obtained. After PCR amplification, these fragments were then cloned into T vectors and were transformed into competent cells subsequently. Ninety-two microsatellite sequences were randomly isolated from 70 positive clones. Twenty-one out of 27 pairs of designed microsatellite primers were screened out from the microsatellite sequences, and 10 out of the 21 microsatellite loci were used to investigate the genetic diversity of three populations of M. fortis, Hunan (wild), Hunan (domesticated), and Ningxia (domesticated). All the 10 microsatellite loci used to analyze the genetic diversity exhibited a good level of polymorphism. The values of observed number of alleles (Na), effective number of alleles (Ne), observed heterozygosity (Ho), expected heterozygosity (He) and polymorphic information content (PIC) were all the highest in the Hunan (wild) population, lower in the Hunan (domesticated) population, and the lowest in the Ningxia (domesticated) population.

Cite this article

NI Li-Ju, DAO Ling-Yun, BAI Xiong, HU Jian-Hua, GAO Cheng, XIE Jian-Yun . Enrichment and screening of the microsatellite markers in Microtus fortis and its preliminary application in genetic diversity research[J]. Hereditas(Beijing), 2011 , 33(9) : 989 -995 . DOI: 10.3724/SP.J.1005.2011.00989

References

[1] 郑智民, 姜志宽, 陈安国. 啮齿动物学. 上海: 上海交通大学出版社, 2008: 172-174.
[2] Weber JL, May PE. Abundant class of human DNA poly-morphisms which can be typed using the polymerase chain reaction. Am J Human Genet, 1989, 44(3): 388-396.
[3] 苏志杰, 俞远京, 周智君, 马亚东, 唐运安. 用数据库搜索法寻找东方田鼠微卫星引物. 中国现代医学杂志, 2008, 18(9): 1212-1214.
[4] Bloor PA, Barker FS, Watts PC, Noyes HA, Kemp SJ. Microsatelliteenriched library construction. http://www. genomics.liv.ac.uk/animal/Protocol1.html.
[5] 谢建云, 邵伟娟, 胡建华, 高诚. 微卫星技术对近交系小鼠遗传质量的分析. 中国比较医学杂志, 2007, 17(9): 511-515.
[6] Botstein D, White RL, Skolnick M, Davis RW. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet, 1980, 32(3): 314-331.
[7] Cheng HH, Levin I, Vallejo RL, Khatib H, Dodgson JB, Crittenden LB, Hillel J. Development of a genetic map of the chicken with markers of high utility. Poult Sci, 1995, 74(11): 1855-1874.
[8] 王登, 施大钊. PCR方法筛选布氏田鼠小片断插入微卫星文库. 草地学报, 2007, 15(3): 278-282.
[9] Kandpal RP, Kandpal G, Weissman SM. Construction of libraries enriched for sequence repeats and jumping clones, and hybridization selection for region-specific markers. Proc Natl Acad Sci USA, 1994, 91(1): 88-92.
[10] Edwards KJ, Barker JH, Daly A, Jones C, Karp A. Microsatellite libraries enriched for several microsatellite sequences in plants. Biotechniques, 1996, 20(5): 758-760.
[11] Fischer D, Bachmann K. Microsatellite enrichment in organisms with larger genomes (Allium cepa L.). Biotechniques, 1998, 24(5): 796-800, 802.
[12] Weber JL. Infornativeness of human (dC-dA)n•(dG-dT)n polymorphisms. Genomics, 1990, 7(4): 524-530.
[13] 李齐发, 赵兴波, 罗晓林, 姚平, 李宁, 田志华, 吴常信, 谢庄. 牦牛基因组微卫星富集文库的构建与分析. 遗传学报, 2004, 31(5): 489-494.
[14] 佟广香, 闫学春, 匡友谊, 梁利群, 孙效文, 王爱民, 王嫣. 马氏珠母贝微卫星快速分离及遗传多样性分析. 海洋学报, 2007, 29(4): 170-176.
[15] 邓玉营. 黄花柳微卫星富集文库构建及标记筛选[学位论文]. 南京: 南京林业大学, 2005.
Outlines

/