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 RTPCR 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.
MyD88RT-PCRgene cloninggene transfectionWestern Blot
NETL
NSTL
万方数据
维普
