龙眼MYB14的生物信息学分析及其基因表达载体构建  

Bioinformatics and Construction of Expression Vector for Longan MYB14

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

机构地区：[1]广西壮族自治区农业科学院/广西作物遗传改良生物技术重点开放实验室,广西南宁530007 [2]广西大学农学院,广西南宁530004 [3]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 [4]广西壮族自治区农业科学院园艺研究所,广西南宁530007

出　　处：《福建农业学报》2020年第9期929-936,共8页Fujian Journal of Agricultural Sciences

基　　金：国家荔枝龙眼产业技术体系建设专项(CARS-33-10),国家荔枝龙眼产业技术体系良种繁育与生产配套技术岗位(CARS-32-04);国家现代农业产业技术体系广西荔枝龙眼创新团队项目(nycytxgxcxtd-02)。

摘　　要：【目的】克隆龙眼MYB-related基因家族中MYB14基因,并对其进行生物信息学分析和基因表达载体的构建,为进一步研究该基因的生物学功能提供基础数据。【方法】通过转录组测序及生物信息学分析得到龙眼MYB14基因全长序列,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,同时构建超表达载体。【结果】MYB14基因属于MYB-related家族的TBP-like亚家族,ORF长度831 bp,编码276个氨基酸,有一个MYB结合位点。该蛋白属于非跨膜亲水蛋白,亚细胞定位预测定位于细胞质,存在57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大麻亲缘关系最近,同时成功构建了植物超表达载体pBI121-MYB14。【结论】本研究克隆得到了龙眼MYB14基因,成功构建表达载体pBI121-MYB14,为该基因功能的研究奠定了坚实的基础。【Objective】MYB14 in Longan was cloned,its bioinformatics studied,and expression vector constructed.【Method】Full-length sequence of MYB14 was obtained by transcriptome sequencing and bioinformatics analysis.Accordingly,specific primers for the cloning were designed,bioinformatics on the open reading frame sequence established,and overexpression vector constructed.【Result】In the TBP-like subfamily of MYB-related family of genes,MYB14 had an831 bp ORF encoding 276 amino acids with one MYB binding site.It was a non-transmembrane hydrophilic protein with 57 amino acid phosphorylation sites in the cytoplasm.Its secondary structure was irregular crimp andα-helix.The predicted secondary and tertiary structures were highly consistent.The phylogenetic tree on the gene of longan showed a homology with that of cannabis.【Conclusion】MYB14 of Longan was successfully cloned with the constructed expression vector,PBI121-MYB14,which provided reference for the further study on the function of MYB14.

关 键 词：龙眼 MYB-related 基因克隆 表达载体构建 

分 类 号：S667.2[农业科学—果树学]