Construction of human FAT10 eukaryotic plasmid and its expression in human embryonic kidney cell line 293T

Objective To construct the human FAT10 eukaryotic plasmid and analyze its expression in human human embryonic kidney cell line 293T.Methods The full-length FAT10 cDNA was obtained by reverse transcription-polymerase chain reaction(RT-PCR) and cloned into pMD18-T simple vector for sequence analysis.The FAT10 gene was subcloned into pEGFP-C1 plasmid.The resulting recombinant vector pcDNA5-FRT-FAT10 was identified by digestion with restriction endonucleases and transfected into 293T cells.The expression of FAT10 protein was examined by semi-quantitative western blot.Results The full-length human FAT10 cDNA was successfully obtained,and the recombinant plasmid pcDNA5-FRT-FAT10 was successfully constructed.After transfection into 293Tcells,western blot analysis showed that FAT10 was highly expressed in 293T cells.Conclusion The FAT10 eukaryotic expression vector pcDNA5-FRT-FAT10 is constructed successfully FAT10 protein express highly in 293T cells.