文献类型：期刊文献

英文题名：Efficient expression and purification of soluble HarpinEa protein by translation initiation region codon optimization

作者：Cai, Zengying[1];Wang, Zhong[2];Yue, Cheng[1];Sun, Aiyou[1];Shen, Yaling[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shandong Shennong Ecol Technol Res Inst Co Ltd, Shanghai Branch, Shanghai 201114, Peoples R China

年份：2021

卷号：188

外文期刊名：PROTEIN EXPRESSION AND PURIFICATION

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000701984100001)】；

基金：This research was financially supported by the 2021 Key R&D Program of Ningxia Hui Autonomous Region, China (Special Project for the Construction of Modern Agricultural Science and Technology Innova-tion Demonstration Zone) .

语种：英文

外文关键词：Harpin(Ea); TIR codon optimization; Protein expression; HR

摘要：Harpin(Ea) protein can stimulate plants to produce defense responses to resist the attack of pathogens, improve plant immune resistance, and promote plant growth. This has extremely high application value in agriculture. To efficiently express soluble Harpin(Ea) protein, in this study, we expressed Harpin(Ea) protein with a 6x His-tag in Escherichia coli BL21 (DE3). Because of the low level of expression of Harpin(Ea) protein in E. coli, three rounds of synonymous codon optimization were performed on the +53 bp of the translation initiation region (TIR) of Harpin(Ea). Soluble Harpin(Ea) protein after optimization accounted for 50.3% of the total soluble cellular protein expressed. After purification using a Ni Bestarose Fast Flow column, the purity of Harpin(Ea) protein exceeded 95%, and the yield reached 227.5 mg/L of culture medium. The purified Harpin(Ea) protein was sensitive to proteases and exhibited thermal stability. It triggered visible hypersensitive responses after being injected into tobacco leaves for 48 h. Plants treated with Harpin(Ea) showed obvious growth-promoting and resistance improving performance. Thus, the use of TIR synonymous codon optimization successfully achieved the economical, efficient, and soluble production of Harpin(Ea) protein.