黄灯笼辣椒CcCAD1基因的克隆与植物表达载体的构建  

Cloning and Plant Expression Vector Construction of CcCAD1 Gene in Capsicum chinense

作　　者：雷欣 居利香 赵成志 童璐 舒黄英 汪志伟 成善汉[1] Lei Xin;Ju Lixiang;Zhao Chengzhi;Tong Lu;Shu Huangying;Wang Zhiwei;Cheng Shanhan(College of Horticulture,Hainan University,Haikou,570228)

机构地区：[1]海南大学园艺学院,海口570228

出　　处：《分子植物育种》2020年第16期5373-5379,共7页Molecular Plant Breeding

基　　金：国家自然科学基金(31760578)资助。

摘　　要：肉桂醇脱氢酶(cinnamyl alcohol dehydrogenase, CAD)在木质素生物合成中有着重要作用,为了探讨辣椒中CAD基因在木质素合成中的作用,本研究利用RT-PCR方法从黄灯笼辣椒幼嫩叶片中克隆得到CAD1基因全长cDNA,命名为CcCAD1。序列分析表明,CcCAD1 cDNA序列全长1 074 bp,编码357个氨基酸。同源比对显示其与番茄CAD1蛋白的一致性高达96.09%。荧光定量PCR检测结果显示,CcCAD1基因在辣椒根、茎、叶、胎座和果肉中的表达量差异显著,其相对表达量为叶>果肉>茎>根>胎座。利用Bam HⅠ和SacⅠ双酶切连接成功构建pBI121-CcCAD1植物表达载体,为CcCAD1基因转化以及后续的功能分析提供依据。Cinnamyl alcohol dehydrogenase(CAD)plays an important role in lignin biosynthesis,in order to exp-lore the regulation of lignin content by CAD gene in pepper.In this study,the full-length cDNA of CAD1 gene was cloned from the tender leaves of Capsicum chinense by RT-PCR and named CcCAD1.Sequence analysis indicated that the full length of CcCAD1 was 1074 bp and encoded 357 amino acids.The homologous alignment showed that the identity of CAD1 protein was as high as 96.09%with the tomato.The results of real-time PCR showed that the expression of CcCAD1 gene in pepper roots,stems,leaves,placenta and pulp was significantly different,and its re-lative expression was leaf>pulp>stem>root>placenta.The pBI121-CcCAD1 plant expression vector was successfu-lly constructed by double digestion with BamHⅠand SacⅠ,which provide basis for CcCAD1 gene transformation and subsequent functional analysis.

关 键 词：黄灯笼辣椒(Capsicum chinense) 肉桂醇脱氢酶 基因克隆 植物表达载体 

分 类 号：S641.3[农业科学—蔬菜学]