华北农学报Issue(1)：73-76,4.DOI:10.7668/hbnxb.2015.01.012
拟南芥抗病相关基因T1N6_22的原核表达分析
Prokaryotic Expression Analysis of Resistance-related Gene T1N6_22 from Arabidopsis thaliana
摘要
Abstract
This study was aimed to construct prokaryotic expression vector of resistance-related gene T1 N6_22 from Arabidopsis thaliana and obtain the purified protein T1N6_22 with high-efficiency expression.The CDS of T1N6_22 was amplified by RT-PCR technology using the cDNA of the Arabidopsis Col-0 and was fused into a pro-karyotic expression vector pGEX 4 T-1 .Restriction enzyme digestion and sequencing showed that the recombinant vector pGEX4T-1-T1N6_22-GST was successfully constructed and transformed into E.coli BL21 cells.The results indicated that the pGEX4T-1-T1N6_22-GST with the predicted molecular weight of about 57 kDa was successfully ex-pressed in E.coli BL21 strain.SDS-PAGE indicated that the best expression quantity was induced with 0.1 mmol/L IPTG treatment for 3 h.These results would provide basis for screening interacting proteins of T 1N6_22 and the reg-ulation mechanism in Arabidopsis resistance .关键词
拟南芥/T1N6_22/原核表达/纯化Key words
Arabidopsis thaliana/T1N6_22/Prokaryotic expression/Purification分类
农业科技引用本文复制引用
蒋琛茜,瓮巧云,樊锦涛,王冠宇,董丽萍,邢继红,董金皋..拟南芥抗病相关基因T1N6_22的原核表达分析[J].华北农学报,2015,(1):73-76,4.基金项目
国家自然科学基金项目（31200203）；河北省自然科学基金项目（C2012204032；C2014405010）；高等学校博士学科点专项科研基金项目 （）
