大豆酪氨酸氨基转移酶基因的克隆与表达分析

Cloning and Expression Analysis of Glycine max Tyrosine Aminotransferase Gene

采用电子克隆与实验克隆相结合的方法获得了大豆酪氨酸氨基转移酶基因的cDNA序列,GenBank登录号为DQ003328。序列分析结果表明,该cDNA序列含有一个编码425个氨基酸的完整的开放读码框,5′非翻译区具有多个同框终止密码子,3′端具有3个加尾信号和polyA尾巴。启动子区除含有通用核心元件外,还含有许多与光反应有关的作用元件。氨基酸序列比对和系统发育分析结果显示,不同物种之间酪氨酸氨基转移酶的氨基酸序列同源性较高。电子表达分析和RT-PCR组织表达分析结果表明,该基因的表达量与组织中叶绿体含量具有很高的关联,强光逆境能够上调该基因的表达。

A cDNA sequence of tyrosine aminotransferase gene was cloned by using in silico cloning combined with experimental RT-PCR with a GenBank accessory number DQ003328.Nucleotide sequence analysis showed that the cDNA had an intact open reading frame（ORF） encoding 425 amino acids.Some same frame stop codons were found in 5′ untranslated region and three tailing signal and a polyA tail were found in 3′ region.Many light responsive elements were found in its promoter region including common core promoter elements.Protein multiple-alignment and phylogenetic analysis suggested that soybean TAT was strong similarity in different plant species.Expression analysis of in silico and RT-PCR results showed that its expression was high correlated with chloroplast content and was up-regulated by high light stress.Conclusively,soybean TAT gene had cloned and its expression was also successfully analyzed by in silico and RT-PCR in this study.