Transcription factor ETS2 regulates genes involved in development, differentiation, angiogenesis, proliferation, and apoptosis. In addition, it is one of the core reprogramming factors responsible for the generation of human cardiomyocytes from adult somatic cells. In this study, we report the heterologous expression of human ETS2 in E. coli to produce a highly pure recombinant protein. To accomplish this, the codon-optimized 1507 bp coding sequence of the human ETS2 gene in fusion with a His-tag, a cell-penetrating peptide, and a nuclear localization sequence was cloned in the protein expression vector and transformed into E. coli strain BL21(DE3) for expression. The recombinant protein was purified to homogeneity under native conditions using immobilized metal ion affinity chromatography, and its identity was confirmed by Western blotting with an ETS2 antibody. Using far-UV circular dichroism spectroscopy, we have demonstrated that the recombinant protein has retained its secondary structure, predominantly comprising of random coils and β-sheets. Prospectively, this biological recombinant ETS2 protein can substitute viral and genetic forms of ETS2 in a cell reprogramming process to facilitate the generation of clinical-grade cells. It can also be used to investigate its molecular role in various biological processes and diseases and for biochemical and structural studies.

转录因子ETS2调节涉及发育，分化，血管生成，增殖和凋亡的基因。另外，它是负责从成年体细胞产生人心肌细胞的核心重编程因子之一。在这项研究中，我们报道了人ETS2在E中的异源表达。大肠杆菌产生高纯度的重组蛋白。为了实现这一点，所述人的密码子优化的1507 bp的编码序列ETS2与His-标签，细胞穿透肽，和核定位序列的融合基因在蛋白表达载体克隆并转化到ë。大肠杆菌用于表达的菌株BL21（DE3）。使用固定的金属离子亲和色谱在自然条件下将重组蛋白纯化至同质，并通过ETS2抗体的Western印迹确认其身份。使用远紫外圆二色光谱，我们已经证明重组蛋白保留了其二级结构，主要由无规卷曲和β-折叠组成。潜在地，这种生物重组ETS2蛋白可以在细胞重编程过程中替代ETS2的病毒和遗传形式，以促进临床级细胞的生成。它也可用于研究其在各种生物过程和疾病中的分子作用以及用于生化和结构研究。