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SP0306蛋白是肺炎链球菌TIGR4菌株中的一种假想的转录因子,但其蛋白三维结构及生物学功能尚未明了,生物信息学分析提示其可能调控碳水化合物代谢相关基因的表达。成功构建了SP0306蛋白的全长表达载体PET28a-sp0306,利用大肠杆菌BL21(DE3)菌株进行原核表达,获得了以可溶形式表达的目的蛋白。经Ni-NTA柱亲和层析及DEAE阴性离子交换层析纯化后,获得了高纯度的目的蛋白。采用悬滴气相扩散法获得了质量较好的SP0306蛋白晶体,并初步进行了晶体X射线衍射,为其最终的三维结构解析及生物学功能研究奠定了基础。
SP0306 protein is an imaginary transcription factor in Streptococcus pneumoniae TIGR4 strain. However, the three-dimensional structure and biological function of SP0306 protein are unknown. Bioinformatics analysis suggests that it may regulate the expression of carbohydrate metabolism-related genes. The full-length expression vector PET28a-sp0306 of SP0306 protein was successfully constructed and prokaryotic expressed in E. coli strain BL21 (DE3) to obtain the target protein in soluble form. Purification by Ni-NTA column affinity chromatography and DEAE negative ion exchange chromatography, the purity of the target protein was obtained. The crystal structure of SP0306 with good quality was obtained by the hanging drop gas diffusion method. The crystal structure of SP0306 was preliminarily analyzed by X-ray diffraction, which laid the foundation for the final three-dimensional structure analysis and biological function study.