HU Ying-kao,MENG Xian-ping,LI Ya-xuan,et al.Cloning and Expression Analysis of Glycine max γ-tocopherol Methyltransferase Gene[J].Soybean Science,2011,30(02):198-204.[doi:10.11861/j.issn.1000-9841.2011.02.0198]
大豆γ-生育酚甲基转移酶基因的克隆与表达分析
- Title:
- Cloning and Expression Analysis of Glycine max γ-tocopherol Methyltransferase Gene
- 文章编号:
- 1000-9841（2011）02-0198-07
- 关键词:
- 大豆; γ-生育酚甲基转移酶; 基因克隆; 电子表达分析
- 分类号:
- S 565.1
- 文献标志码:
- A
- 摘要:
- 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。
- Abstract:
- cDNA sequence of γ-tocopherol methyltransferase gene was cloned by using in silico cloning combined with experimental RT-PCR from Glycine max with a GenBank accession number AY960126. Nucleotide sequence analysis showed that the cDNA has an intact open reading frame (ORF) encoding 350 amino acids. Two same frame stop codons were found in 5′ untranslated region and two tailing signals and a polyA tail were found in 3′ region. Many light responsive elements were found in its promoter region including many common core promoter elements. The deduced protein contained a signal peptide located in chloroplast and two γ-tocopherol methyltransferase motifs. Protein multiple-alignment and phylogenetic analysis suggested that GmTMT was strong similarity in different plant species. Expression analysis with in silico and RT-PCR results showed that its expression was closely correlated with chloroplast content. However, high light stress has no effect on its expression.
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备注/Memo
基金项目：留学回国人员择优资助项目。