| 凡纳滨对虾遗传多样性研究中AFLP体系的建立 |
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| 引用本文： | 王伟毅,徐景祥,阮灵伟.凡纳滨对虾遗传多样性研究中AFLP体系的建立[J].台湾海峡,2012,31(3):440-445. |
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| 作者姓名： | 王伟毅 徐景祥 阮灵伟 |
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| 作者单位： | 1. 国家海洋局第三海洋研究所、国家海洋局海洋生物遗传资源重点实验室,福建厦门,361005
2. 国家海洋局第三海洋研究所、国家海洋局海洋生物遗传资源重点实验室,福建厦门361005;厦门大学生命科学学院,福建厦门361005 |
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| 基金项目： | 国家现代农业产业技术体系建设专项资金资助项目 |
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| 摘 要： | 凡纳滨对虾是当今世界公认的重要对虾养殖品种之一,然而良种选育研究仍相对落后,亟需采用分子遗传学手段加以解决.本文以凡纳滨对虾为材料,对预扩增体系、选择性扩增体系中的稀释倍数、dNTP浓度、Taq浓度、Mg2+浓度、引物浓度进行比较,结果表明,300 ng基因组DNA采用5U PstI和5UMseI双酶切4 h,16℃连接过夜,连接产物稀释10倍进行预扩增,预扩增产物稀释50倍进行选择性扩增,能够得到稳定丰富的条带,最终建立了适用于凡纳滨对虾的AFLP反应体系.本研究又进一步应用优化后的AFLP反应体系,从64对选择性扩增引物组合中筛选出12对多态性高的引物组合,便于对凡纳滨对虾基因多态性位点进行分析,从而为凡纳滨对虾的遗传多样性研究奠定基础.
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| 关 键 词： | 海洋生物学 凡纳滨对虾 遗传多样性 AFLP技术 反应体系 |
Establishment of AFLP fingerprinting in genetic diversity analysis of Litopenaeus vannamei |
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| WANG Wei-yi
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XU Jing-xiang
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RUAN Ling-wei.Establishment of AFLP fingerprinting in genetic diversity analysis of Litopenaeus vannamei[J].Journal of Oceanography In Taiwan Strait,2012,31(3):440-445. |
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| Authors: | WANG Wei-yi XU Jing-xiang RUAN Ling-wei |
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| Institution: | 1(1.Key Laboratory of Marine Biogenetic Resources,Third Institute of Oceanography,SOA,Xiamen 361005,China; 2.College of Life Science,Xiamen University,Xiamen 361005,China) |
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| Abstract: | Although Litopenaeus vannamei is the most economically important shrimp species in the world,broodstock breeding is still limited and molecular genetics seems to be a promising method to overcome the problem.This study is to establish an AFLP reaction system for Litopenaeus vannamei.We analyzed the dilution multiple,the concentration of dNTP,Taq,Mg2＋ and primers for the pre-amplification and selective amplification.It showed that clear and repeatable gel profiles could be obtained under the reaction protocol,with 300 ng genomic DNA digested by 5U of PstI and MseI,then conjugated at 16℃ overnight,the ligation products diluted for 10 times as template for pre-amplification.After pre-amplification,the products were diluted for 50 times and used as template for selective amplification.Using the optimized reaction system,we selected 12 informative primer combinations out of 64,which would be useful for the further study of Litopenaeus vannamei genetic diversity.The establishment of the system might lay a foundation for the application of AFLP techniques for genetic diversity studies of Litopenaeus vannamei. |
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| Keywords: | marine biology Litopemteus vannamei genetic diversity AFLP reaction system |
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