Simultaneous expression of SMYD3 and EGFP in NIH3T3 cells

摘要

SET and MYND domain-containing protein 3 (SMYD3) is a novel histone methyltransferase involved in human carcinogenesis. To establish the transgenic cell model of SMYD3 for the further studies on the characteristics and functions of SMYD3, in the present study, the human SMYD3 gene was subcloned into pIRES2-EGFP eukaryotic expressing vector. The recombinant plasmid was transfected into NIH3T3 cells. The Enhanced Green Fluorescent Protein (EGFP) expression was assayed by fluorescence microscopy. The expression level of SMYD3 was examined by RT-PCR and Western blot. The effects of SMYD3 on proliferation were evaluated by MTT assay. The results showed that the recombinant plasmids were constructed successfully. After transfection, SMYD3 and EGFP could be expressed simultaneously and independently in mice cells, and EGFP fluorescence could reflect the expression of SMYD3. This transgenic cell lines would facilitate to the study of SMYD3 and the screening of novel antitumor drugs.