拟南芥At4g12500基因原核表达载体的构建

摘要

[ Objective] This study aimed to construct the prokaryotic expression vector for Arabidopsis AlAgl2500 gene. [ Method] Genomic DNA of wild-type Arabidopsis Col-0 was extracted as template for amplification of At4gl2500 gene fragment. The target fragment was double-digested with BamH I and Xho I, and ligated with prokaryotic expression vector pET-28a. [Result] Prokaryotic expression vector pET28a-At4gl2500 was successfully constructed and transformed into Escherichia coli expression strain BL21 (DE3). [Conclusion] This study laid the foundation for subsequent expression and functional analysis of At4gl2500 gene.%[目的]构建拟南芥At4g12500基因的原核表达载体.[方法]以野生型Col-0拟南芥基因组DNA为模板,采用PCR技术扩增出At4g12500基因编码序列,用BamH Ⅰ和XhoⅠ对目的片段进行双酶切后,与BamH Ⅰ和Xho Ⅰ双酶切后的pET-28a质粒连接,以得到原核表达载体.[结果]该试验成功地构建了pET28a-At4g12500原核表达载体,并已转化至E.coil BL21( DE3)表达菌株中.[结论]为后续At4g12500基因的表达与功能分析奠定基础.