中国生物工程杂志

Aim: To construct eukaryotic recombinant expression vector pCMV－Myc－fat1 and express fat1 gene in human kidney 293T cells. Methods: The cDNA of fat1 was amplified by RT-PCR from Caenorhabditis elegans. Constructed and identified the vector containing fat1 gene by endonucleases digestion and PCR. Expression plasmids were transfected into 297T cells; the expression of fat1 was detected by Western Blot; the distribution of this gene was analyzed by Indirect Immunofluorescense Assay (IFA). Results: Constructed eukaryotic recombinant expression vector pCMV－Myc－fat1;fat1 gene was effectively expressed in 293T cells and detected by Western Blot and laser scanning confocal microscope. Conclusions: We construct eukaryotic recombinant expression vector pCMV－Myc－fat1 successfully through DNA recombinant technique; The expression of fat1 was detected; the distribution of expressed fat1 was both on the cell membrane and in cytoplasm.