Abstract:

BACKGROUND: Nuclear factor-κB (NF-κB) is a key actor in tumorigenesis, PDLIM2 gene can mediate the termination of NF-κB activation and relieve the apoptosis suppression of NF-κB on tumor cell.
OBJECTIVE: To clone PDLIM2 gene and construct a eukaryotic expression vector pIRES2-EGFP-PDLIM2.
METHODS: PDLIM2 gene, cloned from total of fresh bladder tissue by reverse transcription polymerase chain reaction (RT-PCR), and pIRES2-EGFP plasmid were digested by Bam HI and Xho Ⅰ double endonucleases and linked with each other. The integrity and accuracy of PDLIM2 in pIRES2-EGFP-PDLIM2 recombinant plasmid was indentified by enzyme digestion and sequencing. The expression intensity of GFP report gene transfected by EJ cells were observed under a fluorescence microscope and the PDLIM2 expression level was determined by RT-PCR. 
RESULTS AND CONCLUSION: PDLIM2 gene sequence contained in pIRES2-EGFP-PDLIM2 recombinant plasmid was verified correctly by enzyme digestion as well as sequence analysis. After being transfected into EJ cells, highly efficient expression of PDLIM2 gene contained in pIRES2-EGFP-PDLIM2 recombinant plasmid were detected at mRNA level. PDLIM2 gene can be cloned from bladder tissue by means of RT-PCR. pIRES2-EGFP-PDLIM2 recombinant plasmid has been constructed successfully with high efficient expression in transfected EJ cells.