SUMO修饰IFNγ信号转导域偶联截短型TGF-β II型受体的融合蛋白可溶性表达条件优化  

Optimization of soluble expression of a fusion protein by conjugation of IFNγsignal transduction domain to truncated TGF-βtype II receptor with SUMO modification

作　　者：董溢馨 王小花[2] 刘海峰[1] DONG Yi-xin(College of Life Sciences Mudanjiang Medical University,Mudanjiang 157011,China)

机构地区：[1]牡丹江医学院生命科学学院,黑龙江牡丹江157011 [2]牡丹江医学院基础医学院,黑龙江牡丹江157011

出　　处：《牡丹江医学院学报》2023年第3期14-17,共4页Journal of Mudanjiang Medical University

基　　金：黑龙江省省属高等学校基本科研业务费项目(2022-KYYWF-0698)。

摘　　要：目的优化小分子泛素相关修饰物(SUMO)修饰IFNγ信号转导域偶联截短型TGF-βII型受体的融合蛋白SUMO-BiPPB-mIFNγ-tTRβⅡ可溶性表达的条件。方法将已转化重组质粒pET28a/SUMO-BiPPB-mIFNγ-tTRβⅡ的大肠杆菌Rosetta(DE3)在不同异丙基-β-D-硫代半乳糖苷浓度(0.1、0.3、0.6和1 mmol/L IPTG)、不同诱导温度(37℃、16℃和20℃)和时间(6 h、16 h和20 h)条件下进行诱导,SDS-PAGE分析SUMO-BiPPB-mIFNγ-tTRβⅡ的表达情况。结果在20℃、IPTG为0.6 mmol/L诱导20 h时,重组蛋白SUMO-BiPPB-mIFNγ-tTRβⅡ可溶性表达量最高,约占上清中总蛋白的20.24%。结论获得SUMO-BiPPB-mIFNγ-tTRβⅡ可溶性表达最佳条件,为进一步制备生物活性的蛋白和后续功能研究奠定基础。Objective To optimize the soluble expression of a chimeric protein SUMO-BiPPB-mIFNγ-tTRβⅡ,which is the IFNγsignal transduction domain fused to truncated TGF-βtype II receptor with SUMO modification.Methods The E.coli Rossta(DE3)containing recombinant plasmid pET28a/SUMO-BiPPB-mIFNγ-tTRβⅡwas induced under different IPTG concentrations(0.1,0.3,0.6 and 1 mmol/L),different induction temperatures(37℃,16℃and 20℃)and different induction times(6 h,16 h and 20 h).SDS-PAGE was applied to analyze the expression of SUMO-BiPPB-mIFNγ-tTRβⅡ.Results SUMO-BiPPB-mIFNγ-tTRβⅡwas highly expressed as a soluble form at 20℃and 0.6 mmol/L IPTG for 20 h,which accounts for 20.24%of the total supernatant proteins.Conclusion The optimal conditions for soluble expression of SUMO-BiPPB-mIFNγ-tTRβⅡwere successfully obtained,which provides foundation for preparation of active protein and research on the function.

关 键 词：融合蛋白 可溶性表达 小分子泛素相关修饰物 

分 类 号：R735.7[医药卫生—肿瘤]