杉木Cl4CL基因的克隆及表达分析

Cloning of Cl4CL Gene and Its Expression Analysis in Chinese Fir

4-香豆酸辅酶A连接酶(4CL)催化肉桂酸、对-香豆酸等生成相应的辅酶A脂,是调控木质素单体合成的关键酶。本研究在杉木中克隆了4CL基因,长度为1 596 bp,包含完整开放阅读框,编码531个氨基酸,命名为Cl4CL基因。Cl4CL酶包含20种氨基酸,其中酸性氨基酸含量微高,并含有信号肽,属于分泌蛋白。Cl4CL包含两种高度保守的肽基序,分别为肽基序BoxⅠ(SGTTSRPKGV)和肽基序BoxⅡ(GEVCIRG),其中肽基序BoxⅠ属于AMP结合功能域,肽基序BoxⅡ含有催化反应基团。通过对Cl4CL基因的表达分析发现低氮胁迫下Cl4CL呈现下调表达。

4-coumaric acid coenzyme A ligase(4CL)catalyzes cinnamic acid and para-coumaric acid produces corresponding coenzyme A lipids which is the key enzyme to regulate lignin monomer synthesis.In this study,4CL gene was cloned from Chinese fir.The sequence was 1596 bp in length,and contained a complete open reading frame(ORF),which encoding 531 amino acids and named Cl4CL.The bioinformatics analysis of Cl4CL showed that it contained 20 kinds of amino acids,of which the acid amino acid content was slightly high,and it contained signal peptide,which belonged to secretory protein.Two highly conserved peptide motifs,namely peptide motif BoxⅠ(SGTTSRPKGV)and peptide motif BoxⅡ(GEVCIRG),were obtained by analyzing the Cl4CL functional domain.The peptide motif BoxⅠbelongs to the AMP binding domain,and the peptide motif BoxⅡcontains the group of catalytic reaction.Through the analysis of Cl4CL expression found that under low nitrogen stress was down-regulated.