桃果实PpAO基因的电子克隆及生物信息学分析  

In silico cloning and bioinformatic analysis of PpAO in peach fruit

作　　者：王可[1] 邵兴锋[1] 龚一富[2] 张兴龙[1] 

机构地区：[1]宁波大学食品科学与工程系,浙江宁波315211 [2]宁波大学生物与海洋科学系,浙江宁波315211

出　　处：《食品工业科技》2013年第17期193-196,205,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金(31000825);浙江省自然科学基金(Y3090537)

摘　　要：研究克隆桃果实中的PpAO(Prunus persica ascorbate oxidase)基因,并对其序列进行了生物信息学分析。研究采用电子克隆和RT-PCR(reverse transcription PCR)相结合的方法分离桃果实PpAO基因的cDNA全长序列,利用各种软件对序列进行生物信息学分析。实验成功获得全长1913bp的PpAO基因序列,该序列具有完整的开放阅读框(ORF,open reading frame),编码18个氨基酸的信号肽和518个氨基酸的成熟酶。对ORF进行PCR的验证结果和电子克隆的结果一致。序列分析显示该基因的氨基酸序列同大豆和葡萄具有很高的相似度,含有Cu-oxidase_3,Cuoxidase,Cu-oxidase_2三个同源结构域。The aim of this study was to clone PpAO （ Prunus persica ascorbate oxidase） gene and analyze its transcript levels of peach fruit stored at different low temperatures.This paper isolated PpAO gene complete cDNA length with in silico cloning method and RT-PCR （reverse transcription PCR）from peach fruit, and characterized the sequence in terms of bioinformation by various software packages.One 19]3bp full-length cDNA of PpAO gene was cloned,which codes a complete ORF（open reading frame）including 18 amino acids for signal peptide and 518 amino acids for mature enzyme.The ORF check test was conducted and the result suggested that the sequence of PpAO gene through PCR was nearly the same with that in silico cloning.The amino acid sequence of PpAO gene in peach fruit was very similar to that of Vitis vinifera and Gkycine max, containing 3 conserved domains which were Cu-oxidase_3, Cu-oxidase, Cu-oxidase_2.

关 键 词：桃果实 抗坏血酸氧化酶 电子克隆 

分 类 号：TS255.1[轻工技术与工程—农产品加工及贮藏工程]