马哈利樱桃PGIP cDNA克隆及序列分析

Cloning and sequencing of PGIP cDNA from Prunus mahaleb

以马哈利樱桃 (Prunus mahaleb L .)为材料 ,通过 RT- PCR获得了 1 0 4 5 bp的目的片段 ,经克隆测序 ,证实该片段包含 1个完整的开放阅读框架 ,该阅读框架由 990碱基组成 ,编码 3 3 0个氨基酸。该序列与杏、梨、苹果的 PGIP c DNA序列同源性分别达 97.2 %、83 .4%和83 .6 % ,可能编码的氨基酸与杏、梨、苹果的 PGIP c DNA所编码的氨基酸的同源性分别达到96 .7%、85 .2 %和 85 .2 %。与已经克隆的 PGIP DNA序列的对比分析表明 ,PGIP DNA序列中包含 2个外显子和 1个内含子 ,内含子全长 1 47bp,符合 TG- AG规律 ,2个外显子长度分别为 5 81 bp、46 4

Specific primers,synthesized according to conservative regions of apple,pear and apricot PGIP gene,were used to acquire PGIP cDNA by reverse transcription polymerase chain reaction.A fragment of 1 045 bp was obtained and cloned into pUCm T vector.Sequencing results showed that the fragment was 97.2%?83.4%?83.6% homology to the cDNA sequences of apricot,pear and apple respectively.A open reading frame comprised by 990 bp encoding 330 amino acid was also found.The possible amino acid was 96.7%?85.2%?85.2% homology to that of apricot,pear and apple respectively.Compared to PGIP DNA sequence of PGIP(Genbank accession nuimber AF263465),the PGIP cDNA sequence was spliced by two exons,one is 584 bp and another 416 bp,the deleted intron was 147 bp and was stared with TG and ended with AG as usual.