茶树‘紫娟’肉桂酰辅酶A还原酶基因(CCR)克隆及序列分析

Cloning and Sequence Analysis of Cinnamoyl-CoA Reductase Gene from Tea Plant[Camellia sinensis(L.)O.Kuntz]

通过克隆茶树肉桂酰辅酶A还原酶基因(CCR)的cDNA序列,为研究其蛋白结构和功能奠定基础。对利用cDNA-AFLP技术获得的‘紫娟’茶树成熟叶片上调的差异表达片段TDF,设计特异引物,利用RACE末端扩增技术,分别扩增出5′端和3′端目的片段,测序后进行拼接获得茶树肉桂酰辅酶A还原酶基因(CCR)cDNA全长序列,并进行序列分析。结果表明:克隆的茶树CsCCR基因cDNA全长1 259bp(GenBank登录号为KJ995737),其中开放阅读框957bp。同源比对发现,与蓖麻、丹参、草莓、番茄的CCR蛋白同源性分别为67%、68%、69%和70%。

In order to lay a foundation for the structure and function studies of CsCCRprotein,CsCCR cDNA of tea plant was cloned.The cDNA-AFLP was applied to up-regulated transcript derived fragment（TDF）from mature leaves of tea plant（Camellia sinensis var.assamica,cultivar ‘Zijuan＇）.TDF was used to design specific primers,and then cloning the 5′-and 3′-end sequence by RACE,and fragment was assembled to get completely cDNA.Finally,CsCCRcDNA sequence was analyzed.The results show that the cDNA of CsCCR was 1 259 bp in length（GenBank accession No.KJ995737）,and contains an open reading frame（ORF）of 957 bp and encoding 319 amino acids residues.It was found that the amino acid sequence of CsCCRhad 67%,68%,69% and 70% homology with Ricinus communis,Salvia miltiorrhiza,Fragaria vesca,and Solanum lycopersicum,respectivly.