【摘要】 本文采用RT PCR技术从人外周血PBMC中克隆HLA A2基因的胞外区 ,拼接上依赖BirA酶的可生物素化序列后 ,装入pET 42b (+)高效表达载体进行诱导表达。并对表达产物进行纯化与复性。结果成功地扩增出HLA A2基因并测序证实 ;构建了融合表达载体pET HLA A2 ,并获表达与纯化。为研究在原核系统中表达、纯化与复性及进一步体外构建MHC I类分子四聚体 ,探讨免疫识别机制奠定了基础。更多还原

【Abstract】 The HLA-A2 was amplified from human peripheral blood mononuclear cells by RT-PCR,and added a 15-amino acid substrate peptide for BirA-dependent biotinylation to the COOH-terminus of human lymphocyte antigen(HLA) heavy chain,then cloned into pET-42b(+) vector,getting expressed and purification successfully. High efficient expression of HLA-A2-BSP-his6 fusion protein lays the foundation for further research expression and purification in prokaryotic system and constructing terameric peptide-HLA complexes to explore the mechanism of immune recognition.更多还原