丹参硫氧还蛋白基因SmTrxh 的克隆和生物信息学分析

李松丽, 王喆之*
陕西师范大学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, 西安710062

通信作者:王喆之;E-mail: zzwang@snnu.edu.cn;Tel: 029-85310260

摘 要:

对丹参 EST 数据库进行 BLAST 同源性比对发现, 登录号为 CV165156 的 EST 序列与硫氧还蛋白基因(Trx)有很高的 同源性。进一步用 PCR 方法从丹参基因组水平上克隆到长 1 806 bp 的 DNA 序列(登录号为 FJ217699), 与 cDNA 序列比对 发现, 该基因(SmTrxh)含有 2 个内含子。生物信息学分析表明, SmTrxh 所编码蛋白的分子质量为 13.4 kDa, 理论等电点为5.53, 无信号肽, 属于定位于细胞质中的稳定类蛋白。该蛋白与其他 7 种植物中的 Trx 高度同源, 同源性介于 68%~74% 之 间。实时定量 PCR 检测的结果显示, SmTrxh 在丹参中为组成型表达基因, 在根、茎和叶中都有表达, 主要在根部表达, 茎 中的表达量最低。

关键词:生物信息学; 实时定量PCR; 丹参; 硫氧还蛋白

收稿:2008-12-01   修定:2009-02-24

资助:国家“十一五”科技支撑计划(2006BAI06A12-04)。

Cloning and Bioinformatics Analysis of SmTrxh in Salvia miltiorrhiza Bunge

LI Song-Li, WANG Zhe-Zhi*
Key Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China, Shaanxi Normal University, Xi’an 710062, China

Corresponding author: WANG Zhe-Zhi; E-mail: zzwang@snnu.edu.cn; Tel: 029-85310260

Abstract:

The expressed sequence tags of Salvia miltiorrhiza were analyzed using the BLAST approach of NCBI. One of these sequences (CV165156) showed high homology with thioredoxin gene. A 1 806 bp DNA sequence of the SmTrxh (GenBank accession number FJ217699) was cloned by PCR from genome of S. miltiorrhiza. The sequence aligned with cDNA showed that there were two introns in the encoding region. Bioinformatics analysis showed that the calculated molecular mass was 13.4 kDa and theoretical isoelectric point was 5.53. SmTrxh was a stable protein without signal peptide. Amino acids homology of SmTrxh compared with other seven plants ranged from 68% to 74%. Real-time PCR result showed that the SmTrxh was a constitutive expression gene, which expressed in root, stem and leaf. The mRNA of SmTrxh was most abundant in root, and the least in stem.

Key words: bioinformatics; real-time PCR; Salvia miltiorrhiza; thioredoxin

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