大豆脱氢抗坏血酸还原酶基因的电子克隆及进化分析  

IN SILICO CLONING AND EVOLUTION ANALYSIS OF DEHYDROASCORBATE REDUCTASE cDNA FROM GLYCINE MAX

作　　者：李晓晓[1] 李蕊[1] 李雅轩[1] 蔡民华[1] 胡英考[1] 

机构地区：[1]首都师范大学生命科学学院,北京100037

出　　处：《大豆科学》2007年第1期45-50,共6页Soybean Science

基　　金：北京市教委科技发展计划项目基金资助(200310028112)

摘　　要：利用生物信息学和实验验证的技术路线,以拟南芥脱氢抗坏血酸还原酶基因cDNA序列为信息探针,对大豆EST数据库进行同源搜索,经同源性比对和序列组装,获得全长为955bp的大豆脱氢抗坏血酸还原酶基因的cDNA序列(GenBank登录号为DQ006810),经RT-PCR扩增,分子克隆和序列分析验证,结果表明与电子克隆完全一致。该cDNA序列具有完整的开放阅读框架(ORF,36bp-815bp),推测编码蛋白为259个氨基酸。通过与几种植物脱氢抗坏血酸还原酶蛋白序列的比较,发现该基因具有较高的保守性。结果表明根据物种间同源基因序列,对跨物种间EST数据库进行同源检索筛选,拼接,是新基因克隆的一条有效途径。Using bioinformatics strategy and validating by experiment, a novel dehydroascorbate reductase gene（DHAR） of Glycine max was Cloned and identified which was being blasted by search of Glycine max EST database with homologous gene cDNA of Arabidopsis thaliana. This sequence was confirmed by RT-PCR, molecular cloning and sequencing. The full length Glycine max DHAR gene cDNA of 955bp（GenBank accession,DQ006810） encoded dehydroascorbate re- ductase （DHAR） of 259 amino acid and contained a complete ORF of 777bp. Compared with several plants, we found the gene was almost conserved. The results revealed that it was a convenient technique for cloning novel gene by searching EST database with homologous gene of model plants such as the Arabidopsis thaliana.

关 键 词：大豆 电子克隆 RT-PCR脱氢抗坏血酸还原酶(DHAR) EST 

分 类 号：S565.1[农业科学—作物学]