mpl/G-CSFR融合受体缺失体与突变体的构建

Construction of deletants and mutants of mpl/G-CSFR chimera receptor

目的 :构建mpl/G_CSF融合受体胞内区不同功能域缺失体以及酪氨酸点突变体 ,并获取稳定转染细胞株 ,用以研究受体不同区域以及不同酪氨酸的功能。方法 :用普通PCR法扩增融合受体短截体 ,用重叠延伸PCR法和QickChange定点突变法获得融合受体酪氨酸点突变体 ,并将mpl/G_CSFR全长融合受体转染BaF3细胞。结果与结论 :获得mpl/G_CSF融合受体 3个短截体和 4个点突变体 ,并筛选到全长融合受体的稳定转染细胞株 ,为深入研究受体的功能奠定了基础。

Objective:To construct the deletants and tyrosine dot mutants of chimera receptor according to the different functional motifs in order to study the function of intracellular mpl/G-CSFR chimera receptor, and to select the stable transfected cells.Methods:Normal PCR method was used to get the deletants, overlay extension PCR method and QickChange mutation method were applied to obtain the tyrosine dot mutants, and the liposome was used for transfection.Results and Conclusion:The 3 deletants and 4 tyrosine dot mutants of mpl/G-CSFR chimera receptor were obtained, the full-length chimera receptor was transfected into BaF3 cells, and the stable transfected cells were selected. The experimental results are important for further study on the functions of different motifs of the receptor.