Abstract:Abstract: To construct the eukaryotic expression vector for biotin AP-tagged FOXM1 and purify the biotin-labeled FOXM1 based on the high specificity, high affinity interaction between biotin and streptavidin will provide a foundation for the follow-up identification of FOXM1-interacting proteins. The eukaryotic expression vectors pcDNA3.1-AP-FOXM1 and pcDNA3.1-BirA containing E. coli biotin ligase (BirA) were constructed respectively and co-transfected into HEK 293T cells. The expression of the biotin-labeled FOXM1 in the cells was confirmed by silver nitrate staining and Western-blot with HRP-labeled Streptavidin. The biotinylated FOXM1 was purified with streptavidin agarose beads and the efficiency of the pull-down of FOXM1-interacting proteins were also analyzed. The constructed eukaryotic expression system produced biotinylated FOXM1 in cells. This protein enabled a one-step method for detection and purification of FOXM1 in Western-blot and pull-down experiments without using antibodies. The establishment of eukaryotic expression system of biotinylated FOXM1 provided a technical foundation for the identification of FOXM1-interacting proteins and the study of FOXM1 functions.
引用本文:
邹红春, 谭桂湘, 谭拥军. 生物素化FOXM1真核表达体系的构建及鉴定[J]. 生命科学研究, 2016, 20(1): 16-20.
ZOU Hong-Chun, TAN Gui-Xiang, TAN Yong-Jun. Construction and Identification of Biotinylated FOXM1 Eukaryotic Expression System. Life Science Research, 2016, 20(1): 16-20.