天津农业科学2024，Vol.30Issue(10)：1-7,7.DOI:10.3969/j.issn.1006-6500.2024.10.001
谷子糖基转移酶基因SiDDOST的克隆及表达载体的构建
Cloning of Glycosyltransferase Gene SiDDOST and Construction of Expression Vector in Foxtail Millet
摘要
Abstract
In order to explore the biological function and regulatory role of glycosyltransferase gene in foxtail foxtail foxtail fox Mao Maoliang amplified the 1 338 bp cDNA sequence of the glycosyltransferase gene(SiDDOST)containing the complete coding region,and analyzed the characteristics of the full-length sequence of SiDDOST by bioinformatics methods,and the target gene fragment was double-digested with the restriction enzyme Nco I and Pml I,respectively,and the gene fragment and the vector fragment were recov-ered,and the glycosyltransferase gene fragment was linked to the overexpression vector pCAMBIA3301 by T4DNA ligase,and the re-combinant expression vector was transformed into E.coli DH5α.It was verified by electrophoresis detection of recombinant expression vector and empty vector,PCR of bacterial solution,and double enzyme digestion analysis.The results showed that the CDS region of glycosyltransferase gene was 1 323 bp and encoded 441 amino acids,the range of 30 amino acids at the N-terminus of DDOST pro-tein was poorly conserved,and there were three insertion/deletion mutations in DDOST protein compared with 8 other plant species.This indicated that the gene fragment of interest had been successfully inserted into the expression vector and the transcription direc-tion was correct.In conclusion,the overexpression vector of foxtail millet glycosyltransferase gene constructed in this study lays a foundation for subsequent genetic transformation and functional verification in Arabidopsis thaliana,rice or foxtail millet.关键词
谷子/糖基转移酶基因/克隆/生物信息学分析/载体构建Key words
foxtail millet/glycosyltransferase gene/cloning/bioinformatics analysis/vector construction分类
农业科技引用本文复制引用
李兴杰,王振山,栾孟澳,贾小平..谷子糖基转移酶基因SiDDOST的克隆及表达载体的构建[J].天津农业科学,2024,30(10):1-7,7.基金项目
国家自然科学基金(31471569) （31471569）
