沙棘PEPCK基因的克隆与生物信息学分析

Cloning and Bioinformatics Analysis of PEPCK Gene in Sea Buckthorn

磷酸烯醇式丙酮酸羧激酶(PEPCK)是调节植物糖异生途径的限速酶。本研究从沙棘果实的转录组测序结果中筛选出PEPCK基因序列,利用RT-PCR技术对其进行克隆,利用生物信息学相关方法分析其编码蛋白的理化性质和结构特征。结果表明,该基因开放阅读框(open reading frame, ORF)长度为1 989 bp,编码一个由662个氨基酸组成的亲水性蛋白,主要由α-螺旋(30.21%)、β-折叠(17.22%),无规则卷曲(47.13%)和β-转角(5.44%)组成,其ORF序列与克莱门柚(Citrus clementina)同源性最高,氨基酸序列与川桑(Moras notabilis)和洋蓟(Cynara cardunculus var. scolymus)的有较高的同源性。该结果为进一步研究沙棘PEPCK蛋白功能以及揭示植物糖异生途径的调控机制提供理论基础。

Phosphoenolpyruvate carboxykinase(PEPCK)plays a key role in gluconeogenesis pathway in plant,which involved in the malic acid metabolism.In this study,the PEPCK gene sequence was gained from transcriptome sequencing data of sea buckthorn and specific primers were designed for amplification.Afterwards PEPCK gene was cloned using RT-PCR method and bioinformatics analysis were carried out.The results showed that the ORF(open reading frame)of PEPCK was 1989 bp length,encoding a hydrophilic protein of 662 amino acids.It was mainly composed ofα-helix(30.21%),β-folding(17.22%),irregular curl(47.13%)and a small amount of β-turn(5.44%).Its ORF sequence had close phylogenetic relationships with Citrus clementina and amino acid sequence had close phylogenetic relationships with Moras notabilis and Cynara cardunculus var.scolymus.These results provide a theoretical basis to investigate the regulatory mechanism of malate metabolism in sea buckthorn.