可溶性单链MHC-肽四聚体的构建研究
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Preparing of the single-chain MHC-peptide complex tetramer
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    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-

    Abstract:

    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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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(自然科学版),2009,29(3):328-332

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  • 收稿日期:2008-08-19
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