文章摘要
邵文雨,朱 楠,杨广顺.可溶性单链MHC-肽四聚体的构建研究[J].南京医科大学学报,2009,29(3):328~332
可溶性单链MHC-肽四聚体的构建研究
Preparing of the single-chain MHC-peptide complex tetramer
投稿时间:2008-08-19  
DOI:10.7655
中文关键词: 细胞毒性T淋巴细胞  四聚体  表位  重折叠
英文关键词: CTL  tetramers  epitope  refold
基金项目:
作者单位
邵文雨 第二军医大学附属东方肝胆外科医院胆道二科,上海 200438 
朱 楠  
杨广顺  
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中文摘要:
      目的:构建检测抗原特异性细胞毒性T淋巴细胞的单链MHC-肽四聚体?方法:通过在MHC重链基因中引入生物素化位点,并在轻链和重链基因中引入同一限制性酶切位点连接MHC轻链和重链构建可生物素化的单链MHC分子?结果:重组子以包涵体形式表达得到单链MHC分子,重折叠?生物素化?荧光标记后得到活性单链MHC-肽四聚体?结论:单链MHC分子操作简便,提高了四聚体构建效率?
英文摘要:
      Objective:To construct a single chain MHC-epitope tetramers for detecting epitope specific cytotoxic T lymphocyte. Motheds:The C terminus of β2m fusing with extracellular domains of HLA A0201 was connected with a accessional BirA substrate peptide by a linker through restriction enzyme digestion and ligation. Results:Expression of the recombinant in a pET systerm as inclusion body was followed by refolded in the presence of the antigenic peptide. After biotinylization of the MHC-peptide complex, the tetramers was obtained by the coupling of a fluorchrome conjugated streptavidin with the complex. Conclusion:The novel method of constitution of tetramers was more efficient than the old labor-consuming one.
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