Construction of eukaryotic expression vector of MyD88 gene and detection of its expression

AIM: To construct eukaryotic expression vector of the gene of myeloid differentiation factor 88(MyD88) and to detect its expression in GES-1 cells.METHODS: Total RNA was extracted from the mononuclear cells of human peripheral blood.The full length cDNA of MyD88 gene was amplified by RT-PCR and then cloned into pcDNA3.1 /myc-His(-)A vector by Kpn I and Nhe I restriction enzyme sites.The gene was identified and confirmed by sequencing and then was transfected into GES-1 cells.The expressed MyD88 protein was detected by Western blot analysis.RESULTS: The sequencing results confirmed that the eukaryotic expression vector of MyD88 was correctly constructed and the results of Western blot showed its good expression in GES-1 cells.CONCLUSION: pcDNA3.1/myc-His(-) A-MyD88 has been successfully constructed and expressed in GES-1 cells,which provides a foundation for further investigation of the molecular constitution and function of MyD88.