丹参普遍胁迫蛋白基因(SmUSP)的克隆及表达模式分析

邝静, 生华, 武玉翠, 葛茜, 张媛, 王喆之*
药用资源与天然药物化学教育部重点实验室, 西北濒危药材资源开发国家工程实验室, 陕西师范大学生命科学学院, 西安 710062

通信作者:王喆之;E-mail: zzwang@snnu.edu.cn;Tel: 029-85301260

摘 要:

通过BLAST比对分析丹参EST数据库, 发现一个与普遍胁迫蛋白(USP)基因家族同源性较高的基因序列, 命名为SmUSP, GenBank登录号为JX416284。依据该序列, 从DNA和cDNA水平克隆得到该基因全长, 经分析发现该基因无内含子序列, 包含一个长711 bp的完整开放读码框(ORF), 编码236个氨基酸残基。生物信息学分析显示, SmUSP编码蛋白的相对分子量25.5 kDa, 为无信号肽和跨膜结构域的疏水酸性蛋白; SmUSP具有UspA结构域和ATP结合位点, 属于USP蛋白UspFG亚家族。实时荧光定量PCR分析表明, SmUSP在丹参的根、茎、叶、花中均有表达, 在根中的表达量较高。同时, SmUSP表达受茉莉酸甲酯(MeJA)、水杨酸(SA)、脱落酸(ABA)以及脱水胁迫的上调诱导, 推测该基因可能在丹参防御反应中发挥作用。

关键词:SmUSP; 生物信息学分析; 表达模式分析

收稿:2012-12-03   修定:2013-03-12

资助:陕西省自然科学基础研究计划项目(2012JQ4013)、中央高校基本科研业务费专项资金资助(GK201102017)、陕西教育学院自然基金(2012KJ044)。

Cloning and Expression Analysis of Universal Stress Protein Gene (SmUSP) from Salvia miltiorrhiza Bunge

KUANG Jing, SHENG Hua, WU Yu-Cui, GE Qian, ZHANG Yuan, WANG Zhe-Zhi*
Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education; National Engineer-ing Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China; College of Life Sciences, Shaanxi Normal University, Xi’an 710062, China

Corresponding author: WANG Zhe-Zhi; E-mail: zzwang@snnu.edu.cn; Tel: 029-85301260

Abstract:

A sequence showed high homology with universal stress proteins (USPs) was found by BLAST searching in EST database of Salvia miltiorrhiza. The full-length cDNA and gDNA sequences of the gene were cloned and named as SmUSP. The GenBank accession number is JX416284. SmUSP contained an intact open reading frame (ORF) of 711 bp without intron and encoded 236-amino acid peptides. Bioinformatic analysis showed SmUSP was a stable acidic protein with a predicted molecular weight of 25.5 kDa. There is no signal peptides and transmembrane domain in SmUSP. SmUSP protein has an UspA domain with an ATP-binding site and can be classified into the UspFG subfamily of USP. The results of real-time quantitative PCR revealed that SmUSP was expressed in roots, stems, leaves and flowers of S. miltiorrhiza, and most abundant in roots. Up-regulated expression of SmUSP could be induced by methyl jasmonate (MeJA), salicylic acid (SA), abscisic acid (ABA) and drought, indicating that SmUSP might be involved in plant defenses.

Key words: SmUSP; bioinformatics analysis; expression analysis

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