山东医药Issue(31)：5-7,3.DOI:10.3969/j.issn.1002-266X.2015.31.002
mM-CSF 及其剪切体重组逆转录病毒表达载体的构建
Construction of membrane-bound macrophage colony-stimulating factor and recombinant retroviral expression vector of spliceosome
摘要
Abstract
Objective To construct membrane-bound macrophage colony-stimulating factor ( mM-CSF) and recombi-nant retroviral expression vector of brachytmema mutation of 30 amino acide located in the intracellular region of mM-CSF ( mM-CSF-Δ) .Methods The retroviral vectors MSCV-PGK-GFP-mM-CSF and MSCV-PGK-GFP-mM-CSF-Δwere con-structed and identified by DNA recombinant techniques.Recombinant and empty vectors were used to transfect the packa-ging Phoenix cells.HEK293 cells were infected by the viral supernatants.After being sorted by flow cytometry, three posi-tive cell lines were obtained.Results HEK293 cells were successfully infected by retroviruses in packaging Phoenix cells and control retrovirus.Stable expressing cell lines, HEK293-M and HEK293-M-Δas well as control cell line HEK293-V, were established.The expression of mM-SCF and mM-CSF-Δwas detected by RT-PCR and Western blotting in HEK293 cells and its membrane protein expression was also detected by flow cytometry.Conclusion The mM-CSF and recombinant retroviral expression vector of spliceosome were successfully constructed.关键词
mM-CSF/剪切体/逆转录病毒表达载体/HEK293细胞系Key words
membrane-bound macrophage colony-stimulating factor/spliceosome/retroviral expression vector/HEK293 cell line分类
医药卫生引用本文复制引用
马翠花,廖金凤,王大刚,刘淑艳,任倩,郑国光..mM-CSF 及其剪切体重组逆转录病毒表达载体的构建[J].山东医药,2015,(31):5-7,3.基金项目
国家自然科学基金资助项目（81370634）。 （）
