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云南松基因组微卫星富集文库的构建

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Abstract:

采用磁珠富集法构建云南松微卫星富集文库。云南松基因组DNA经RsaⅠ酶切,与特定接头连接,再用接头特异#br#引物进行PCR扩增。连接扩增产物与用生物素标记的(AG)12、(AT)12、(CG)12、(GT)12、(ACG)12、(ACT)12和(CCA)8#br#探针杂交,通过链霉亲和素偶联的磁珠捕捉含接头和微卫星序列的片段并扩增,将获得的片段连接到pMD-19T载体上,转化至大#br#肠杆菌JM109感受态细胞中,成功构建了云南松微卫星富集文库。通过PCR检测从文库中筛选阳性克隆,在383富集阳性菌落中#br#获得阳性克隆257个,经测序分析,在获得的159条序列中,有143条含有SSR,其中完美型占65.73%,非完美型占23.78%,混#br#合型占10.49%。结果表明,磁珠富集法构建云南松基因组微卫星文库高效、可行的,文库的构建为微卫星位点的分离、遗传多样#br#性的分析等奠定基础。

References

[1]  中国科学院中国植物志编辑委员会. 中国植物志[M]. 第7 卷. 北京: 科学出版社, 1978: 211-282. [2] 金振洲, 彭鉴. 云南松[M]. 昆明: 云南科技出版社, 2004: 6,21, 332. [3] Sehgal D, Raina SN. DNA markers and germplasm resource diagnostics: new perspectives in crop improvement and conservation strategies[M] //Arya ID, Arya S. Utilization of biotechnology in plant sciences. Meerut, India: Arya Rastogi Press, 2009: 39-54. [4] Varshney RK, Graner A, Sorrells ME. Genic microsatellite markers in plants: features and applications[J]. Trends Biotechnol, 2005,23(1): 48-55. [5] Kalia RK, Rai MK, Kalia S, et al. Microsatellite markers: an overview of the recent progress in plants[J]. Euphytica, 2011, 177(3): 309-334. [6] Weber JL, May PE. Abundant class of human DNA polymorphisms which can be typed using the polymerase chain reaction[J]. The American Journal of Human Genetics, 1989, 44(3): 388-396. [7] Powell W, Machray GC, Provan J. Polymorphism revealed by simple sequence repeats[J]. Trends in Plant Science, 1996, 1(7): 215-222. [8] Glenn TC, Schable NA. Isolating microsatellite DNA loci[J].Methods in Enzymology, 2005, 395: 202-222. [9] 邓欣, 陈信波, 龙松华, 等. 用磁珠富集法分离亚麻基因组微卫 星分子标记[J]. 作物学报, 2008, 34(12): 2099-2105. [10] Weber JL. Informativeness of human(dC-dA)n?(dG-dT)n polymorphisms[J]. Genomics, 1990, 7(4): 524-530. [11] Williams JG, Kubelik AR, Livak KJ, et al. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers[J].Nucleic Acids Research, 1990, 18(22): 6531-6535. [12] Wu K, Jones R, Danneberger L, Scolnik PA. Detection of microsatellite polymorphisms without cloning[J]. Nucleic Acids Research, 1994, 22: 3257-3258. [13] C if a rel l i RA , G all ite l li M, Ce l lin i F. R a n dom a m pl i fie d hybridization microsatellites(RAHM): isolation of a new class of microsatellite-containing DNA clones[J]. Nucleic Acids Research, 1995, 23(18): 3802-3803. [14] Richardson T, Cato S, Ramser J, Ket al. Hybridization of microsatellites to RAPD: a new source of polymorphic markers[J].Nucleic Acids Research, 1995, 23(18), 3798-3799. [15] Ender A, Schwenk K, Stadler T, et al. RAPD identification of microsatellites in Daphnia[J]. Molecular Ecology, 1996, 5(3): 437- 441. [16] Lunt DH, Hutchinson WF, Carvalho GR. An efficient method for PCR-based isolation of microsatellite arrays(PIMA)[J]. Molecular Ecology, 1999, 8: 891-894. 99 2014年第1期许玉兰等: 云南松基因组微卫星富集文库的构建 [17] Zane L, Bargelloni L, Patarnello T. Strategies for microsatellite isolation: a review[J]. Mol Ecol, 2002, 11: 1-16. [18] Ostrander EA, Jong PM, Rine J, Duyk G. Construction of smallinsert genomic DNA libraries highly enriched for microsatellite repeat sequences[J]. Proceedings of the National Academy of Sciences of the USA, 1992, 89(8): 3419-3423. [19] Paetkau D. Microsatellites obtained using strand extension: An enrichment protocol[J]. Biotechniques, 1999, 26(4), 690692,694-697. [20] Gao GQ, He GH, Li YR. Microsatellite enrichment from AFLP fragments by magnetic beads[J]. Acta Botanica Sinica, 2003, 45 (11): 1266-1269. [21] Winter AK, Fredholm M, Thomsen PD. Variable n. (dG-dT)(dC-dA) n sequences in the porcine genome[J]. Genomics, 1992, 12(2): 281-288. [22] Refseth UH, Fangan BM, Jakobsen KS. Hybridization capture of microsatellite directly from genomic DNA[J]. Electrophoresis,1997, 18: 1519-1523. [23] Kandpal RP, Kandpal G, Weissman SM. Construction of libraries enriched for sequence repeats and jumping clones,and hybridization selection for region-specific markers[J].Proceedings of the National Academy of Sciences of the USA, 1994,91(1): 88-92.

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