【摘要】 共用 5 2 0个 10碱基随机引物对小麦抗条锈基因Yr5的近等基因系进行了RAPD分析 ,发现了 3个特异性DNA片段S14 96 76 1 、S14 5 3880 、S14 181 950 与Yr5基因连锁 ,其中S14 96 76 1 与Yr5基因紧密连锁 ,遗传距离为 2 7cM。经对特异性DNA片段S14 96 76 1 进行克隆、测序 ,设计了PCR扩增用专化引物SC S14 96 76 1 a和SC S14 96 76 1 b ,用该引物可扩增出与原RAPD引物扩增出的相似的特异DNA片段。由于该引物还可扩增出迁移率极为相近的另 1条非特异带 ,在琼脂糖凝胶上难以分辨 ,需用聚丙烯酰胺凝胶电泳结合银染进行检测。经用F2 分离群体及部分相关品种材料检测 ,已证明该标记的可靠性更多还原

【Abstract】 A total of 520 10 mer random primers were used to identify the RAPD markers linked to the Yr5 gene between the near isogenic line Yr5/6 ×Avocet S and recurrent parent Avocet S. Three polymorphic DNA fragments,S1496 761 ,S1453 880 and S1418 1950 ,were found linked to the Yr5 gene. In which the genetic distance between S1496 761 and Yr5 gene was 2.7 cM. The fragment S1496 761 was recovered from the gel and cloned and sequenced. A pair of specific PCR primers was designed based on the sequence. The specific primers amplified the same fragment about 761bp as the random primer S1496 did. Because the primers could amplify another non specific fragment,the PCR products must be analyzed by electrophoresis on polyacrylamide gels.更多还原