杏RAPD两种凝胶电泳指纹特点及PCR扩增片段的序列分析

为更好地将RAPD技术应用于杏品种资源鉴定及遗传基础的研究,文章首次在选用11碱基长度引物的优化技术体系基础上,分别使用两种凝胶对16个杏品种RAPD的PCR扩增产物进行电泳检测与DNA指纹比较。结果表明,聚丙烯酰胺凝胶电泳检测出的谱带总数量以及多态性谱带的数量均高于琼脂糖凝胶,前者不存在或很少存在共迁移性的现象。根据两种电泳系统获得的标记信息对16个杏品种的遗传关系的分析发现,两者聚类结果存在一定差异。以此,提出了科学利用两种电泳的RAPD标记技术的策略。对随机挑选的24个片段进行克隆与测序,结果发现24个片段都是对应引物的RAPD扩增产物。在不同的11条序列中有3条是编码蛋白的基因片段,说明了RAPD不仅扩增基因组上的非编码蛋白序列,同时可以扩增编码蛋白的基因片段,能够较全面客观地反映不同杏品种间遗传上的差异。

Characterization of Apricot RAPD Fingerprints Using Two Different Electrophoresis and Sequences of PCR Amplified Products

In order to utilize RAPD better in the identification of apricot cultivars and apricot genetic analysis,the RAPD fingerprints,generated by two kinds of electrophoreses,of the genomic DNA of 16 apricot cultivars were compared,and the results showed that the polyacrylamide gel（PAGE） electrophoresis RAPD could present more bands,and more polymorphic bands could be uncovered.Two clustering charts deriving from the data collected from the two kinds of DNA fingerprints were constructed,between which there were some difference.Based on the results mentioned above,we put forward the brief strategy of the use of RAPD in apricot genetic study.The random sequencing of 24 fragments showed that all the fragments were of the RAPD PCR products generated from the corresponding primer,among which four sequences were regions of encoding protein genes,suggesting RAPD can amplify both protein-encoding gene and non-protein encoding sequences and uncover the genetic difference of different apricot cultivars with un-bias DNA information.