辣椒尿黑酸植基转移酶基因(HPT)的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of Homogentisate Phytyltransferase(HPT) Gene from Pepper

作　　者：郭娟[1] 佘明明 游文慧 彭秋菊 蒲玲 申娅琴 Guo Juan;She Mingming;You Wenhui;Peng Qiuju;Pu Ling;Shen Yaqin(School of Life Sciences, Guizhou Normal University, Guiyang,550025)

机构地区：[1]贵州师范大学生命科学学院,贵阳550025

出　　处：《分子植物育种》2019年第8期2448-2457,共10页Molecular Plant Breeding

基　　金：贵州省科学技术基金(黔科合J字LKS[2012]21号);贵州师范大学博士科研项目;中国科学院2013年度"西部之光"人才培养项目共同资助

摘　　要：尿黑酸植基转移酶(HPT)是控制植物中维生素E合成的一个关键限速酶,本实验利用RACE方法成功克隆辣椒尿黑酸植基转移酶基因并对其进行生物信息学分析,结果表明,HPT开放阅读框长为1 227 bp,共编码408个氨基酸,属于异戊烯转移酶PT_UbiA超家族。理化性质分析显示HPT蛋白分子量为4.61 kD,半衰期为30 h,理论等电点为9.52,含35个磷酸化位点,无信号肽,是一个疏水性蛋白质,存在9个跨膜结构域,亚细胞定位在类囊体膜上。二级结构预测结果表明HPT蛋白α-螺旋占59.07%、延伸链占10.54%、无规则卷曲占30.39%。进化树分析显示辣椒HPT与番茄HPT同源性较高,为今后利用辣椒尿黑酸植基转移酶通过基因工程技术提高辣椒维生素E的含量提供了重要依据。Homogentisate phytyltransferase(HPT) is a key rate-limiting enzyme for vitamin E biosynthesis in pants.In this study, the HPT gene from pepper was successfully cloned by RACE method, and the bioinformatics analysis of it was carried out. The results indicated that the open reading frame of HPT was 1 227 bp, encoding 408 amino acids,which belonged to the PT_UbiA superfamily. The physicochemical properties analysis showed that the molecular weight of HPT protein was 4.61 kD, the half-life was 30 h and the theoretical isoelectric point was 9.52. There were35 phosphorylation sites in amino acid and no signal peptide. It was a hydrophobic protein and had nine transmembrane domains, with the subcellular localization on thylakoid membrane. Prediction results of secondary structure suggested that alpha-helix, elongation chain, random coil in HPT protein accounted for 59.07%, 10.54%,30.39%, respectively. The phylogenetic tree analysis showed the pepper HPT had higher similarity with tomato HPT,which could provide an important basis for using the HPT to improve the content of vitamin E in pepper by genetic engineering technology in the future.

关 键 词：辣椒 维生素E 尿黑酸植基转移酶(HPT) 基因克隆 生物信息学 

分 类 号：S641.3[农业科学—蔬菜学]