葎草DNA的提取及AFLP标记反应体系的建立  

DNA Extraction and Establishment of AFLP Marker Reaction System of Humulus scandens

作　　者：肖理慧[1] 高武军[1] 张小莉[2] 卢龙斗[1] 邓传良[1] 路淑霞[1] 

机构地区：[1]河南师范大学生命科学学院,新乡453007 [2]河南大学生命科学学院,开封475001

出　　处：《分子植物育种》2008年第2期393-398,共6页Molecular Plant Breeding

基　　金：河南省教育厅自然科学基金(2006180014);新乡市科技攻关项目(07N050)资助

摘　　要：本文对CTAB法、改良CTAB法和SDS法提取雌雄异株植物葎草(Humulus scandensL.)基因组的方法进行了比较分析,并在此基础上对影响葎草AFLP扩增的关键因素模板浓度、酶切时间、选择性扩增模板浓度进行了优化。结果表明,改良的CTAB法更适合葎草基因组DNA的提取;同时利用AFLP分子标记技术,采用EcoI-MseⅠ酶切组合,共筛选27对引物组合,确定了适用于葎草AFLP反应的最佳酶切连接、预扩和选扩体系。同时发现2对引物组合E-ACG+M-CTA和E-AAC+M-CAC共扩增出5条雌雄性别特异条带。The optimized AFLP （amplified dragment length polymorphism） analysis system was established by comparing and analysing the methods of CTAB （eetyl rrimethyl ammonium bomide）, improved CTAB and SDS （sodium dodeeyl sulfate） used for genomie DNA extraction of female and male leaves of a dioecious Humulus scandens and by studying the essential factors （i.e. the density of the genomie DNA template, the time of the re- striction reaction and the density of the selective expanding template） that influenced the results of AFLP method. The results indicated that the method of improved CTAB should be more appropriate for Humulus scandens ge- nomie DNA extraction. By the AFLP technique, using MseI-EcoRI enzyme-cutting combination, the optimal restrietion-ligase reaction system, pre-expanding system and selective expanding system were determined and two pairs of primers （i.e. E-ACG＋M-CTA and E-AAC＋M-CAC） with five distinctive AFLP sex-markers were selected from 27 pairs of primer combination.

关 键 词：葎草 DNA提取 AFLP反应体系的建立 

分 类 号：Q943.2[生物学—植物学]