HLA-B* 2705-β_2m-多肽复合物的制备及纯化

Preparation and purification of HLA-B* 2705-β_2m-peptide complex

目的:制备并纯化HLA-B*2705-β2微球蛋白(β2-microglobulin,β2m)-多肽复合物。方法:将pET21 a-HLA-B*2705和pET21 a-β2m质粒在宿主菌E.coliBL21(DE3)中诱导表达,高效表达的HLA-B*2705蛋白与β2m,在HLA-B*2705限制性抗原肽〔来源于人类免疫缺陷病毒(H IV)gp120蛋白的九肽NH2-GRAFVTIGK-COOH〕存在的情况下,通过稀释复性折叠成HLA-B*2705-β2m-多肽复合物单体,经Western印迹鉴定,分子筛纯化。结果:成功地制备出HLA-B*2705-β2m-多肽复合物。结论:HLA-B*2705-β2m-多肽复合物的成功制备及纯化,为进一步研究HLA-B*2705与NK细胞Ig样受体(killer Ig-like receptor,KIR)之间的相互作用奠定了基础。

Objective： To prepare and purify HLA-B ＊ 2705-β2m-peptide complex. Methods： pET21a-HLA-B ＊ 2705 And pET21a-β2m were transformed into E. eoli BL21, respectively, and their expression was induced by IPTG. The protein expressed was purified and refolded in vitro by limiting dilution with HLA-B ＊ 2705, β2m and HLA-2705 binding peptide （NH2-GRAFVTIGK-COOH from the gp120 protein of HIV） to produce the HLA-B ＊ 2705-β2m-peptide monomer. The product was detected by Western blot. Results： The experimental results showed that the protein of HLA-B ＊ 2705-β2m- peptide complex was successfully constructed and refolded. Conclusion：The highly efficient expression of this protein lays the foundation for the further studies on exploration of the mechanism of immunization recognition between HLA-B ＊ 2705 and killer Ig-like receptor of natural killer cell.