李RAPD扩增片段的不同电泳检测及其序列特征分析  

Research on the Different Eelectrophoretic Detection of Plum RAPD Amplified Fragments and Analysis of Its Sequence

作　　者：初建青[1] 张敬茹[2] 杨光[1] 谭洪花[1] 房经贵[1] 

机构地区：[1]南京农业大学园艺学院,江苏南京210095 [2]中国农业科学院果树研究所,辽宁兴城125100

出　　处：《安徽农业科学》2010年第25期13633-13636,13639,共5页Journal of Anhui Agricultural Sciences

基　　金：2008年江苏省"青蓝工程"人才培育计划

摘　　要：[目的]为更好地将随机扩增多态(RAPD)技术应用于李品种资源鉴定及遗传基础的研究提供理论依据。[方法]以16个李品种为材料,分别使用琼脂糖凝胶和聚丙烯酰胺凝胶(PAGE)对RAPD的PCR扩增产物进行电泳检测,并对检测效果进行比较。[结果]PAGE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。根据2种电泳系统获得的标记信息构建了2张树状聚类图,2张聚类图的聚类情况基本一致。通过对随机挑选的23个片段进行克隆与测序,结果发现23个片段都是对应引物的RAPD扩增产物。其中有4条是编码蛋白的基因片段,说明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。[结论]RAPD不仅能扩增基因组上的非蛋白编码序列,而且还可以扩增编码蛋白的基因片段,是能够较全面反映基因组序列信息理想的DNA标记技术。[Objective]The theoretical basis of the better application of the random amplified polymorphic（RAPD） technology in identification of plum germplasm resource and the research of its genetic basis was provided.[Method] 16 plum cultivars being taken as the experimental materials,the PCR amplification products of RAPD were detected with the agarose gel and polyacrylamide gel （PAGE） electrophoresis and the testing results were compared.[Results] the Total number of the band and the polymorphic band in the method of PAGE electrophoresis detection were many more than these in the method of agarose gel electrophoresis detection.According to two kinds of information from the electrophoresis detection system,the clustering figure was constructed and there was almost consistent situation in the two figures.It was found these 23 fragments,which were cloned and sequenced after randomly selected,were the products from their corresponding RAPD primer amplification.Among them,four fragments were encoded protein gene,indicating not only the sequence of non-coding protein in genome but also the fragment of protein-coding gene could be amplified by the method of RAPD.[Conclusion] Both the sequence of non-coding protein in genome and the fragment of protein-coding gene could be amplified by the method of RAPD,which was a desired marker technique being able to more fully reflect the genomic DNA sequence information.

关 键 词：李 RAPO 琼脂糖凝胶电泳 PAGE 

分 类 号：S662.3[农业科学—果树学]