普通烟草鸟氨酸转氨酶基因Ntδ-OAT的克隆与表达分析  

Cloning and Expression Analysis of Orn-δ-aminotransferase Gene Ntδ-OAT in Nicotiana tabacum

作　　者：焦蓉[1,2] 刘贯山[1] 刘好宝[1] 王树林[1,2] 侯娜[1,2] 王全贞[1,2] 靳义荣[1,2] 刘朝科 冯祥国 胡晓明 

机构地区：[1]中国农业科学院烟草研究所/农业部烟草类作物质量控制重点开放实验室,青岛266101 [2]中国农业科学院研究生院,北京100081 [3]川渝中烟工业公司,成都610000

出　　处：《植物遗传资源学报》2011年第4期588-593,600,共7页Journal of Plant Genetic Resources

基　　金：国家烟草专卖局重点科技项目(110200701021)

摘　　要：鸟氨酸转氨酶是以鸟氨酸为前体合成脯氨酸的关键酶,对植物适应逆境胁迫起关键作用。本研究根据其已知保守序列设计兼并引物,从普通烟草品种中烟14中克隆到一条425bp的中间片段,应用RACE方法设计特异性引物扩增得到5′末端和3′末端cDNA序列,经BLAST验证,首次从普通烟草中克隆到了鸟氨酸转氨酶基因,命名为Ntδ-OAT,其GenBank登录号为GU144571。该基因cDNA全长为1781bp,共编码477个氨基酸。系统进化树分析显示,Ntδ-OAT基因的进化符合传统的生物学分类,主要功能域在生物进化中保守性较高。Real-time PCR分析表明,Ntδ-OAT基因受干旱、高盐、低温、ABA等多种胁迫处理的诱导表达,可能参与普通烟草体内抗渗透胁迫过程。Orn-δ-aminotransferase is the key enzyme for Pro synthesis from Orn,so that it plays an important role in adaptation to environmental stress in plants.A 425-bp segment was cloned from tobacco（Nicotiana tabacum,zhongyan 14）using degenerate primer designed from the known conserved regions of Orn-δ-aminotransferase gene.Special primers derived from the 425-bp segment were used to amplify the 5′ and 3′ ends of the cDNA by RACE.Comparison and analysis of the sequence was performed with BLAST which proved that it was the first time to get the clone of Orn-δ-aminotransferase gene from tobacco.The full-length cDNA was termed Ntδ-OAT（GenBank accession GU144571）which was 1781 bp in length and encoded a protein of 477 amino acids.According to phylogenetic tree,the evolution of Ntδ-OAT corresponds with traditional biological classification.Its functional domain has a high conservative property in the process of biological evolution.By the analysis of real-time PCR,It was determined that Ntδ-OAT could express under many inducements such as drought,high salt,low temperature,ABA and so on,from which we might conclude that Ntδ-OAT plays a role in the resistance to osmoticstress.

关 键 词：普通烟草 鸟氨酸转氨酶 Nt-δOAT 克隆 表达分析 

分 类 号：S572[农业科学—烟草工业]