[Construct Mxi1 mutation gene expression vector from leukemia cell line KG1].

OBJECTIVE

To construct the eukaryotic expression vector for Max interacting protein 1 (Mxi1).

METHODS

The full length cDNA of Mxi1 gene obtained from fetal lymphocyte and KG1 cells were inserted into plasmid pDs-red2-N1 respectively to generate pDs-red2-N1/Mri1 (wild/mutation type). Then the recombinant vector was transfected into Cos-7 cells via liposome. 48 hours post transfection, mRNA of Mri1 gene was detected by RT-PCR and Mxi1 protein expression was detected by flow cytometry and fluorescence microscope in the Cos-7 cells.

RESULTS

The eukaryotic expression vector of Mxi1 was constructed and transfected into eukaryotic cells successfully. The expression of red fluorescence protein in the transfected Cos-7 cells was observed under fluorescence microscope which implied the expression of Mxi1. The transfect efficiency of both wild and mutation type were in a high level in 3 days after transfected, which lasted to 6 d. RT-PCR amplified the total RNA extracted from the transfected Cos-7 cells could find increased mRNA level of Mxi1 gene.

CONCLUSION

We successfully constructed the eukaryote expression vector for Mri1 gene; Cos-7 cells transfected with the vector via liposome could express Mxi1 protein. These could be useful for the further study of the Myc gene modulation.