目的为获得能在酵母表面正确折叠的人衰变加速因子。方法通过 PCR技术从 DAF- p Bluescript M13- (Amp+ )质粒扩增出全长的 DAF c DNA,酶切后克隆到酵母表面呈现载体 p YD1,构建 DAF- p YD1重组质粒 ,转化酵母细胞 ,流式细胞仪检测所呈现的 DAF。结果通过流式细胞仪检测到酵母细胞表面有 DAF分子的表达 ,并且此表面呈现的 DAF与针对DAF不同表位的 3株单抗均能结合。结论酵母表面呈现的 DAF保持了天然 DAF分子的构象 ,为进一步构建 DAF的突变体库奠定了基础 The goal is to obtain accelerated human decay accelerators that fold correctly on yeast surfaces. Methods The full-length DAF cDNA was amplified by PCR from the DAF-p Bluescript M13- (Amp +) plasmid. After digested, the full-length DAF cDNA was cloned into the yeast surface to construct the vector pYD1. The recombinant DAF-p YD1 plasmid was constructed and transformed into yeast Cytometry detects the presence of DAF. Results The expression of DAF was detected on the surface of yeast cells by flow cytometry, and the surface DAF was able to bind to the three McAbs against different epitopes of DAF. Conclusions The DAF presented on the surface of yeast maintains the conformation of native DAF molecule, which lays the foundation for further construction of the mutant library of DAF