Construction and expression of an eukaryotic expression plasmid of Mdfic with c-Myc tag.

Objective To construct an eukaryotic expression plasmid carrying the novel transcriptional factor Mdfic with c-Myc tag. Methods The full length cDNA sequence of Mdfic was amplified by PCR. Annealed oligonucleotides containing a c-Myc antibody recognition sequence and an in-frame stop codon were cloned downstream of the Mdfic cDNA sequence and then subcloned into the pcDNA3.1 (+) expression vector to construct the eukaryotic expression plasmid pcDNA-Mdfic-Myc. The recombined plasmid was sequenced and then transfected into C2C12 cell by lipofectamineTM 2000. The expression of Mdfic-Myc was confirmed by RT-PCR and Western blotting assays. Results The eukaryotic expression plasmid pcDNA-Mdfic-Myc was constructed successfully. Results from RT-PCR and Western blot indicated that the Mdfic-Myc fusion protein was expressed successfully in C2C12 cell. Conclusion The recombinant eukaryotic expression pcDNA-Mdfic-Myc plasmid can be used in the functional study of Mdfic and its interacting proteins.