重组斑马鱼 p8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p8蛋白并纯化。方法：PCR扩增斑马鱼p8蛋白基因编码区，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pET-28a-p8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＋柱纯化重组蛋白。结果：构建了pET-28a-p8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12．8×10^3。结论：获得了斑马鱼p8融合蛋白，为其生物学功能研究奠定了基础。

Prokaryotic Expression and Purification of Recombinant p8 Protein of Zebrafish

Objective： To express and purify the recombinant p8 protein of zebrafish in E.coli BL21（DE3）. Meth- ods： The coding sequence of p8 protein of zebrafish was amplified by PCR and inserted into the prokaryotic expression vector pET-28a with 6×His tag to construct the recombinant plasmid pET-28a-pS. The recombinant plas- mid was transformed into E.coli BL21（DE3）, and the expression of fusion protein was induced by IPTG. Ni2＋ met- al chelating column was utilized for the purification of the fusion protein after the expression conditions were opti- mized. Results： Recombinant plasmid pET-28a-p8 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 12.8 kDa expectedly. Conclusion： The fusion p8 protein of zebrafish was successful- ly expressed and purified, which lays the foundation of further study on its biological function.