黄灯笼辣椒CcCASP1基因的克隆与VIGS载体构建  

Molecular Cloning and VIGS Expression Vector Construction of CcCASP1 Gene in Capsicum chinense

作　　者：申龙斌[1] 秦于玲[1] 刘子记[1] 曹振木[1] Shen Longbin;Qin Yuling;Liu Ziji;Cao Zhenmu(Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China(Ministry of Agriculture),Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Sciences,Danzhou,571737)

机构地区：[1]中国热带农业科学院热带作物品种资源研究所,农业部华南作物基因资源与种质创制重点开放实验室,詹州571737

出　　处：《分子植物育种》2022年第20期6742-6749,共8页Molecular Plant Breeding

基　　金：海南省自然科学基金高层次人才项目(2019RC312);中国热带农业科学院基本科研业务费专项(16300320-20030)共同资助。

摘　　要：为研究海南黄灯笼辣椒凯氏带膜蛋白在抗病过程中的功能,利用PCR技术从辣椒叶片cDNA中扩增出CASP基因,对其蛋白序列进行生物信息学分析,并构建病毒诱导基因沉默(VIGS)载体,验证其效果。结果表明:成功克隆CASP基因,并命名为CcCASP1基因。该基因全长为501 bp,编码166个氨基酸。经生物信息学分析表明,CcCASP1蛋白是一个小分子碱性疏水稳定的膜蛋白,存在4个相对保守的跨膜螺旋结构域。系统进化关系分析结果表明,CcCASP1蛋白与番茄和烟草CASP蛋白亲缘关系较近。利用酶切连接的方法构建了VIGS载体,在海南黄灯笼辣椒中成功地沉默了CcCASP1基因,且效果显著,为深入研究Cc CASP1基因在黄灯笼辣椒中的抗病功能提供技术支持。To investigate the function of a casparian strip membrane domain protein of Capsicum chinense Jacquinin during the process of disease resistance,a casparian strip gene was amplified from pepper leaf cDNA by PCR,and its protein sequence was analyzed by bioinformatics.At the same time,the VIGS vector of Cc CASP1gene was constructed,and then the effect of gene silencing was verified.The results showed that a CASP gene was successfully cloned,and name CcCASP1.The full length of Cc CASP1 was 501 bp,encoding 166 amino acids.CcCASP1 is a small molecule alkaline hydrophobic and stable membrane protein with four relatively conserved transmembrane helix domains by protein sequence analysis.The genetic evolutionary relationship analysis showed that CcCASP1 was closely related to tomato and tobacco CASP proteins.The VIGS vector was successfully constructed by enzyme digestion and ligation,and the effect of gene silencing of Cc CASP1 was significant.These results will be helpful to furthermore research the function of CcCASP1 gene during disease response in Capsicum chinense Jacquinin.

关 键 词：黄灯笼辣椒(Capsicum chinense Jacquin) CcCASP1 基因克隆 生物信息学分析 VIGS载体 

分 类 号：S641.3[农业科学—蔬菜学]