龙眼DlZAT10基因克隆与植物超表达载体构建  

Cloning of Longan DlZAT10 Gene and Construction of PlantOverexpression Vector

作　　者：李琳 丁峰[1,2,4] 潘介春 彭宏祥[3] 张树伟[3] 何新华[1,4] 徐炯志[1] 黄幸 王金英[1] 王颖[1] 李浩然 LI Lin;DING Feng;PAN Jiechun;PENG Hongxiang;ZHANG Shuwei;HE Xinhua;XU Jiongzhi;HUANG Xing;WANG Jinying;WANG Ying;LI Haoran(College of Agriculture,Guangxi University,Nanning,Guangxi 530004,China;Guangxi Key Open Laboratory of Crop Genetic Improvement Biotechnology,Nanning,Guangxi 530007,China;Institute of Horticulture,Guangxi Academy of Agricultural Sciences,Nanning,Guangxi 530007,China;State Key Laboratory of Conservation and Utilization of Subtropical Agricultural Biological Resources,Nanning,Guangxi 530004,China)

机构地区：[1]广西大学农学院,广西南宁530004 [2]广西作物遗传改良生物技术重点开放实验室,广西南宁530007 [3]广西壮族自治区农业科学院园艺研究所,广西南宁530007 [4]亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004

出　　处：《热带作物学报》2021年第1期33-39,共7页Chinese Journal of Tropical Crops

基　　金：国家荔枝龙眼产业技术体系项目(No.CARS-33-10);国家现代农业产业技术体系广西荔枝龙眼创新团队项目(No.nycytxgxcxtd-02)。

摘　　要：以龙眼成熟叶片为材料,通过转录组测序及生物信息学分析得到龙眼ZAT10基因全长序列,命名为DlZAT10(GenBank登陆号:MT117769),进一步设计特异引物对其开放阅读框(ORF)全长序列进行克隆和生物信息学分析。DlZAT10基因属于C2H2锌指蛋白家族的C1-2i亚家族,ORF长度为738 bp,编码245个氨基酸,无内含子,有2个典型的C2H2结合位点。该蛋白属于非跨膜亲水蛋白,预测定位于细胞核,存在40个氨基酸磷酸化位点。其二级结构主要由无规则卷曲,以及α-螺旋和β-折叠构成,二级结构与三级结构预测结果高度一致。同源基因进化树分析表明,该基因与柑橘和阿月浑子亲缘关系最近。同时成功构建了植物超表达载体PBI121-DlZAT10,为DlZAT10基因功能的深入研究奠定了基础。Using the mature leaves of longan as the materials,the full-length sequence of the ZAT10,named DlZAT10(GenBankID:MT117769)in the longan transcriptome was obtained by transcriptome sequencing and bioinformatics analysis.Furthermore,specific primers were designed for cloning and bioinformatics analysis of the full-length sequence of its open reading frame(ORF).DlZAT10 belonged to the C1-2i subfamily of the C2H2 zinc finger protein family.The ORF sequence was 738 bp in length,encoded 245 amino acids with two typical C2H2 binding sites of C2H2.This protein was non-transmembrane hydrophilic and predicted to be located in the nucleus with 40 amino acid phosphorylation sites.Its secondary structure was mainly composed of random coils,as well asα-helix andβ-sheet.The predicted results of the secondary structure and tertiary structure were highly consistent.Phylogenetic tree analysis of the homologous gene indicated that the gene was most closely related to citrus and pistachio.At the same time,the plant overexpression vector pBI121-DlZAT10 was successfully constructed,which would provide basis for the in-depth study of the function of DlZAT10 gene.

关 键 词：龙眼 DlZAT10 基因克隆 胁迫 

分 类 号：S667.2[农业科学—果树学]