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采用电子克隆与实验克隆相结合的方法获得了大豆γ-生育酚甲基转移酶基因的cDNA序列,GenBank登录号为AY960126。序列分析结果表明,该cDNA序列含有1个编码350个氨基酸的完整的开放读码框,5′非翻译区具有2个同框终止密码子,3′端具有2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。编码的蛋白质序列含有1个信号肽和γ-生育酚甲基转移酶的特征基序,该蛋白定位于叶绿体中。氨基酸序列比对和系统发育分析结果显示,不同物种之间γ-生育酚甲基转移酶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,但强光逆境对该基因的表达无影响。
The cDNA sequence of soybean γ-tocopherol methyltransferase gene was obtained by a combination of electronic cloning and experimental cloning. The GenBank accession number is AY960126. Sequence analysis showed that the cDNA sequence contained a complete open reading frame (ORF) of 350 amino acids with two stop codons in the 5 ’untranslated region and two tailed tail signals and polyA tail at the 3’ end. In addition to the promoter region contains a common core elements, but also contains many light-response-related elements. The encoded protein sequence contains one signal peptide and the characteristic motif of gamma-tocopheryl methyltransferase, which is localized in the chloroplast. Amino acid sequence alignment and phylogenetic analysis showed that the homology of amino acid sequences of γ-tocopherol methyltransferase among different species was high. The results of electronic expression analysis and RT-PCR showed that there was a high correlation between the expression level of the gene and the content of chloroplast in the tissue, but the light stress had no effect on the expression of the gene.