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丹参硫氧还蛋白基因SmTrxh的克隆和生物信息学分析
引用本文:李松丽,王喆之. 丹参硫氧还蛋白基因SmTrxh的克隆和生物信息学分析[J]. 植物生理学通讯, 2009, 0(5): 433-438
作者姓名:李松丽  王喆之
作者单位: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
基金项目:国家“十一五”科技支撑计划(2006BA106A12-04).
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1806bp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13.4kDa,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

Cloning and Bioinformatics Analysis of SmTrxh in Salvia miltiorrhiza Bunge
Affiliation: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)
Abstract:The expressed sequence tags of Salvia miltiorrhiza were analyzed using the BLAST approach of NCBI. One of these sequences (CV 165156) 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 com- pared 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.
Keywords:bioinformatics  real-time PCR  Salvia miltiorrhiza  thioredoxin
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