尾叶桉GLU4无性系肉桂酰乙醇脱氢酶基因克隆及序列分析  

Cloning and Prokaryotic Expression of Cinnamyl Alcohol Dehydrogenase of Eucalyptus urophylla Clone GLU4

作　　者：陈博雯[1] 蒋华 覃鹏飞 姜伟[4] 

机构地区：[1]广西优良用材林资源培育重点实验室,广西林业科学研究院,广西南宁530002 [2]广西国有维都林场,广西来宾546100 [3]广西国有大桂山林场,广西贺州542800 [4]广西大学生命科学与技术学院,广西南宁530005

出　　处：《西部林业科学》2014年第2期19-24,共6页Journal of West China Forestry Science

基　　金：广西优良用材林资源培育重点实验室开放课题基金项目(12A0302);国家自然科学基金项目(31100473)

摘　　要：根据植物中CAD基因保守序列设计引物,以尾叶桉GLU4嫩茎为材料克隆到其CAD基因,命名为EuCAD。序列分析结果表明,该基因gDNA长2 200 bp,cDNA长1 102 bp,开放阅读区编码356个氨基酸。EuCAD编码的蛋白质序列分析显示,序列中具有完整CAD1保守结构域,并包含NADP结合位点以及2个Zn结合位点。同源性分析结果显示,EuCAD核酸序列与其他5种桉属植物CAD基因同源性达到97%以上,EuCAD编码蛋白与碧桃等植物中CAD蛋白同源性也在79%以上。对EuCAD编码蛋白进行理化性质及结构的生物信息学分析结果显示,该蛋白为一种无信号肽、无跨膜区的酸性蛋白质,二级结构包含典型的α-螺旋和β-折叠,三级结构同源建模显示与拟南芥的AtCAD4同源性极高。系统进化树分析结果显示,该蛋白与桉属、杨属植物亲缘关系较近,与杉科植物较远。A CAD gene was cloned from the young stem of Eucalyptus urophylla clone GLU4 and named EuCAD by using specific primers based on the highly conserved sequences of plant CAD. Sequence analysis indicated that its gDNA and cDNA sequence were 2 200 bp and 1 102 bp respectively, which open reading frame encoding 356 amino acid residues. EuCAD encoding sequence was analyzed, and the result showed it has an entire CAD1 domain, a NADP binding site and two Zn binding sites. The results of homology analysis showed that EuCAD nucleotide sequence had more than 97 % sequence homology with that of other 5 Eucalyptus, and EuCAD encoding protein had more than 79 % homology with that of Prunus persica and others. The physicochemical property and structure of EuCAD encoding protein were analyzed using bioinformatics tools. The result showed it was an acidic protein without signal peptide or transmembrane domain, and its secondary structure has typical α-helix and β-sheet. Tertiary structure modeling showed it was most similar to AtCAD4 in Arabidopsis thaliana. Phylogenetic analysis showed that it has close relative relation with Eucalyptus and Populus, but distant relation with Taxodiaceae.

关 键 词：尾叶桉 肉桂酰乙醇脱氢酶 木质素合成 基因克隆 序列 生物信息学分析 

分 类 号：S792.39[农业科学—林木遗传育种]