中国林学(英文版)2007，Vol.9Issue(3)：208-212,5.
High-level expression of 4-coumarate:coenzyme A ligase gene Pt4CL1 of Populus tomentosa in E. coli
High-level expression of 4-coumarate:coenzyme A ligase gene Pt4CL1 of Populus tomentosa in E. coli
摘要
Abstract
In order to investigate the enzymatic properties of the 4CL1 of Populus tomentosa, the recombinant expression vector pQE31-4CL1 was constructed. The recombinant was identified by three restriction endonucleases, then the vector pQE31-4CL1 was transformed into expression host M15 (pREP4) and induced by isopropyl-α-D-thiogalactoside (IPTG) to express 60 kD fused protein pression temperature declined from 37 to 28℃. The 6×His tag facilitates affinity binding to Ni2+-nitrolotriacetic acid (NTA) and enables one-step purification to acquire the molecular SDS-PAGE electrophoresis purity of the active 4CL1 protein by agarose coupled with Ni2+-NTA affinity chromatography. The optimal substrate for Pt4CL1 was 4-coumarate.关键词
4-coumarate:coenzyme A ligase, Populus tomentosa, prokaryotic expression, enzyme activityKey words
4-coumarate:coenzyme A ligase, Populus tomentosa, prokaryotic expression, enzyme activity分类
农业科技引用本文复制引用
Fan Bing-you,Lu Hai,Jiang Xiang-ning..High-level expression of 4-coumarate:coenzyme A ligase gene Pt4CL1 of Populus tomentosa in E. coli[J].中国林学(英文版),2007,9(3):208-212,5.基金项目
This study was financially supported by the Major State Basic Research Development Program of China (973 Program) (G1999016005). （973 Program）
