Cloning of A Full-Length cDNA Encoding 4-Coumarate:CoA Ligase of Amorpha fruticosa by PCR-Based Methods
法克隆紫穗槐4-香豆酸:CoA连接酶全长cDNA(英文)作者机构：大连理工大学生物工程系大连116029 宝生物大连有限公司大连116600
出 版 物：《Forestry Studies in China》 (中国林学（英文版）)
年 卷 期：2002年第4卷第1期
页 面：13-17页
学科分类：0710[理学-生物学] 071001[理学-植物学] 07[理学]
主 题：gene cloning random amplification of cDNA end PCR Amorpha fruticosa 4 coumarate:CoA ligase lignin
摘 要：An Amorpha fruticosa cDNA encoding 4 coumarate:CoA ligase (4CL), a key enzyme of phenylpropanoid metabolism related to lignin forming, was cloned by degenerating oligo primed polymerase chain reaction (PCR) and rapid amplification of cDNA end (RACE) PCR. We designed 5′RACE primers based on 4CLA1 fragment which obtained from degenerate PCR. Inverse PCR and nested PCR enabled cloning of the remainder fragments of the gene included 5′ and 3′ end sequence. The ORF encodes a polypeptide of 540 amino acids. The predicted amino acid sequence exhibits significant homology with those of other cloned 4CL genes, contain domains typical of predicted 4CL proteins, in particular a postulated AMP binding site, catalytic domain, and conserved Cys residues.
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