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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 different functional domain deletions and tyrosine point mutants in the intracellular domain of mpl / G_CSF fusion receptor and to obtain stable transfected cell lines to study the function of different regions and different tyrosine receptors. Methods: The fused receptor short truncates were amplified by the ordinary PCR method. The fusion mutant tyrosine point mutants were obtained by overlap extension PCR and QickChange site-directed mutagenesis. The full-length mpl / G_CSFR fusion protein was transfected into BaF3 cells . RESULTS AND CONCLUSION: Obtaining 3 short-stranded and 4-point mutant of mpl / G_CSF fusion receptor and screening the stable transfected cell line with full-length fusion receptor laid the foundation for further study on the function of receptor.