AFLP分子标记技术的改进  

Improvement of Amplified Fragment Length Polymorphism (AFLP) Technique

作　　者：刘平武[1] 李赟[1] 蔡强[1] 宋传宇[1] 何庆彪[1] 杨光圣[1] 

机构地区：[1]华中农业大学作物遗传改良国家重点实验室

出　　处：《生物技术通报》2006年第C00期255-259,271,共6页Biotechnology Bulletin

基　　金：国家973项目(2001CB108807);国家863重大专项(2002AA207009);长江学者和创新团队发展计划资助(IRT0442);华中农业大学科研启动项目

摘　　要：对AFLP技术的限制性内切酶组合进行了改进。以两个低频酶组合代替传统的高频酶与低频酶组合进行分子标记筛选，结果表明，采用低频酶组合（EcoRⅠ＋PstⅠ）产生的条带比传统酶切组合（EcoRⅠ＋MseⅠ和PstⅠ＋MseⅠ）要少且分布不均匀，多集中在150～800bp之间，条带信号强度要大，多态性明显得到提高，分子标记筛选成功率也得到了提高，而且其成本更低的特点有利于AFLP技术在我国的进一步推广普及。In this study, AFLP （amplified fragment length polymorphism） was improved on the combination of restriction enzymes by using two average cutting frequency restriction enzymes （EcoR Ⅰ ＋ Pst Ⅰ） to take place the combination of an average cutting frequency and a higher cutting frequency restriction enzyme （ EcoRⅠ ＋ MseⅠ and PstⅠ ＋ MseⅠ） in a single digestion. The results showed that the combination of EeoR Ⅰ ＋ Pst Ⅰ could generate fewer fragments with larger size （most of bands distribute among 150 bp to 800bp） as well as a higher percentage of polymorphism than EcoR Ⅰ ＋ Mse Ⅰ and Pst Ⅰ ＋ Mse Ⅰ . The signal strength and polymorphism of PCR products were improved too. Furthermore, the results described here have great commercial potential because the average cutting frequency enzymes EcoRⅠ and PstⅠ are much cheaper than the high cutting frequency enzyme MseI. In this way, the cost of AFLP analysis can be reduced.

关 键 词：AFLP分子标记 限制性内切酶组合 改进 遗传多样性 

分 类 号：Q78[生物学—分子生物学]