Zhong Ming, Niu Yong-Chun, Xu Shi-Chang, Wu Li-Ren
Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, China.
Yi Chuan Xue Bao. 2002;29(8):719-22.
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 x 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.
总共使用了520个10聚体随机引物来鉴定与近等基因系Yr5/6×阿沃塞特S和轮回亲本阿沃塞特S之间的Yr5基因连锁的RAPD标记。发现三个多态性DNA片段S1496(761)、S1453(880)和S1418(1950)与Yr5基因连锁。其中S1496(761)与Yr5基因之间的遗传距离为2.7厘摩。片段S1496(761)从凝胶中回收、克隆并测序。根据该序列设计了一对特异性PCR引物。特异性引物扩增出与随机引物S1496相同的约761bp片段。由于引物还能扩增出另一个非特异性片段,因此PCR产物必须通过聚丙烯酰胺凝胶电泳进行分析。