| 鸭微卫星标记的筛选及其遗传多样性分析 |
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| 引用本文： | 黄守婷,叶炎,何桂娟,杨建敏,张金耀,雷祖建,肖天放.鸭微卫星标记的筛选及其遗传多样性分析[J].广西农业生物科学,2014(5):986-991. |
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| 作者姓名： | 黄守婷 叶炎 何桂娟 杨建敏 张金耀 雷祖建 肖天放 |
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| 作者单位： | 福建农林大学动物科学学院,福州350002 |
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| 基金项目： | 福建省高等学校科技项目（JA09082）; 福建省高效产学合作科技重大项目（2012N5001）共同资助;感谢福建省龙岩市山麻鸭保种场和福建省石狮市水禽保种中心提供的实验材料. |
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| 摘 要： | 采用磁珠富集法从AFLP片段中筛选微卫星标记,并对5个福建地方鸭品种的遗传多样性进行检测。基因组DNA用EcoRⅠ/MseⅠ内切酶酶切的同时与接头连接,酶切连接产物与用生物素标记的探针杂交,应用磁珠捕获100~2000bp含有微卫星序列的DNA片段并通过pGEM-T载体转化到大肠杆菌DH5a感受态细胞中,构建富集微卫星序列的基因组文库。随机挑选46个阳性克隆,测序获得14条含有微卫星的DNA序列并递交到GenBank（登录号：FJ599499~FJ599512）。设计合成14对微卫星引物,通过PCR优化从中选择5对引物用于5个福建地方鸭品种的遗传多样性分析。结果显示,5对微卫星引物共检测到31个等位基因,各微卫星基因座的有效等位基因数为1.969 7~2.834 4,多态信息含量和杂合度的平均值分别为0.5133和0.7480,遗传多样性丰富,说明磁珠富集法适合用于鸭微卫星标记的分离与筛选,筛选得到的5个微卫星位点可作为有效的遗传标记用于福建省地方鸭品种遗传多样性的研究。
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| 关 键 词： | 鸭 磁珠富集 微卫星 遗传多样性 |
Screening of Microsatellite Molecular Markers and Genetic Diversity Anal- ysis for Duck Breeds |
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| Huang Shouting Ye Yan He Guijuan Yang Jianmin Zhang Jinyao Lei Zujian Xiao Tianfang.Screening of Microsatellite Molecular Markers and Genetic Diversity Anal- ysis for Duck Breeds[J].Journal of Guangxi Agricultural and Biological Science,2014(5):986-991. |
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| Authors: | Huang Shouting Ye Yan He Guijuan Yang Jianmin Zhang Jinyao Lei Zujian Xiao Tianfang |
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| Institution: | Huang Shouting Ye Yan He Guijuan Yang Jianmin Zhang Jinyao Lei Zujian Xiao Tianfang( College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou, 350002) |
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| Abstract: | Microsatellite-enriched libraries of five indigenous duck breeds in Fujian were constructed using an en- richment method with biotin-labeled probes and magnetic beads. Microsatellite markers were selected from AFLP fragments to detect the genetic diversity of the five indigenous duck breeds. Adapters were ligated with genomic DNA fragments digested by EcoR I and Mse I, the products of ligation were hybridized with biotinylated probe, and the 100-2 000 bp fragments containing microsatellite markers were captured by magnetic beads. Then the cap- tured fragments were transformed into DH5ot competent cells by pGEM-T vector, and being sequenced. Finally, 14 microsatellite sequences were randomly isolated from 46 positive clones, and which were submitted to GenBank （accession number： FJ599499-FJ599512）. Fourteen pairs of primers with microsatellite repeats were designed, and 5 microsatellite loci were isolated by PCR optimization, which were used to investigate the genetic diversity of the 5 Fujian indigenous ducks. A total of 31 alleles were detected by the 5 microsatellite loci. The number of effective alleles per microsatellite loci ranged from 1.969 7 to 2.834 4, the mean PIC of 5 microsatellite loci in 5 indigenous duck breeds was 0.513 3, the mean heterozygosity was 0.748 0, showing rich polymorphism. This study demon- strated that the microsatellite-enriched method was a useful technique of microsatellite isolation for the duck breeds. 5 microsatellite makers would provide a tool for assessing polymorphism analysis. |
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| Keywords: | Duck Enrichment by magnetic beads Microsatellite Genetic diversity |
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