重组斑马鱼CD36蛋白的原核表达及纯化

目的：在大肠杆菌中重组表达斑马鱼CD36蛋白胞外区38～432氨基酸残基段并纯化。方法：PCR扩增斑马鱼CD36蛋白的基因编码区，连接到带有6~His标签的原核表达载体pET-28a中，构建重组表达质粒pET28a-CD36，并转化大肠杆菌BL21（DE3），用IPTG诱导表达，优化表达条件后用Ni^2＋柱进行纯化。结果：构建了pET28a-CD36重组质粒；目的蛋白在大肠杆菌中获得表达，亲和纯化后，SDS-PAGE显示相对分子质量为预期的46．8×10^3。结论：获得了斑马鱼CD36融合蛋白，为其生物学功能研究奠定了基础。

Prokaryotic Expression and Purification of Recombinant Zebrafish CD36 Protein

Objective： To construct prokaryotic expression vector of recombinant zebra.fish CD36 protein, and to purify the protein of extracellular amino acid residues 38 to 432 segments. Methods： The coding sequence of zebrafish CD36 protein was amplified by PCR and inserted into the prokaryotic expression vector pET-28a with 6× His tag to construct the recombinant plasmid pET28a-CD36. The recombinant plasmid was transformed into E.coli BL21（DE3）, and the expression of fusion protein was induced by IPTG. Ni2. metal chelating column was utilized for the purification of the fusion protein after the expression conditions were optimized. Results： Recombinant plasmid pET28a-CD36 was constructed and the recombinant protein was expressed in E.coli successfully. After being purified by affinity chromatography, SDS-PAGE showed a clear protein band with a relative molecular weight of 46.8 kD expectedly. Conclusion： The fusion protein of zebrafish CD36 was successfully expressed and purified,which lays the foundation of further study on its biological function.