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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (24): 553-558.

• 水产 渔业 • 上一篇    下一篇

山女鳟（Oncorhynchus masou masou）AFLP体系建立的研究

张 超1,2,佟广香1,匡友谊1,张春雷1,3,尹家胜1,贾钟贺1   

  • 收稿日期:2009-07-10 修回日期:2009-08-11 出版日期:2009-12-20 发布日期:2009-12-20
  • 基金资助:

    科技部支撑计划;农业部水产生物技术重点开放实验室基金

Establishment of AFLP Analysis System for Oncorhyrchus masou masou

  • Received:2009-07-10 Revised:2009-08-11 Online:2009-12-20 Published:2009-12-20

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摘要：

山女鳟（Oncorhynchus masou masou）是一种优良的冷水性养殖鱼类，为了进行山女鳟养殖群体的遗传管理，本研究构建了山女鳟的AFLP分析体系，试验采用两组酶切组合酶切山女鳟基因组DNA，并对酶量及酶切时间、预扩增模板用量、预扩产物稀释倍数进行了优化。结果表明，100ng DNA用3U的EcoRI在37℃酶切3h后，再分别用3U的TaqI和Tru9I在65℃酶切4h连接效果最佳，取3μL连接产物进行预扩增，将预扩增产物稀释50倍进行选择性扩增，可以得到清晰的扩增图谱；对两组酶切组合进行对比发现，EcoRI-Tru9I组合扩增的条带数多于EcoRI-TaqI组合，但后者的多态性位点比例高于前者。

关键词: 大量种子, 大量种子, 加热超干燥, 种子活力

Abstract:

AFLP(Amplified Fragment Length Polymorphism) analysis system for Oncorhyrchus masou masou were established in this study, the genomic DNA were digested by two groups of restriction enzymes, with the relative processes being optimized, including the quantity of the restricting enzymes, the time of digested reaction, quantity of the template for pre-amplification and times of pre-amplification products dilution. It was proved that the results were best with 100ng DNA digested for 3h at 37℃ by 3U EcoRI and then 4h at 65℃ by TaqI and Tru9I respectively, then ligated by ligase. Quite clearly results could be achieved by 3μL ligation products as the template for pre-amplification which products were diluted 50 times for selective amplification. The comparison of the two pairs showed that the group of EcoRI-Tru9I obtained more amplification bands but lower ratio of polymorphic loci than EcoRI-TaqI.