Objective：To construct recombinant adenovirus vector containing PAR-4 gene，so as to lay a foundation for further study of the function of PAR-4 and to illuminate the effect of PAR-4 on islet β cell line-NIT-1. Methods：Oligo Designer 3.0 was used to de－sign the PCR primers：PRKC apoptosis WT1 regulator（pawr）. Then，the gene fragments of pawr were obtained by RT-PCR and insert－ed into the ADV1 vector by restriction enzyme digestion.The recombinant vector plasmid pAdtrack-pawr-CMV was constructed，and then，the recombinant pAdTrack-pawr-CMV was linearized and translated into the competent bacteria with pAd-Easy-1 for homolo－gous recombination to obtain pAdTrack-PAR-4. NIT-1 cell was divided into 4 groups：low glucose control，low glucose+overexpression of PAR-4，high glucose control，high glucose + overexpression of PAR-4，and protein expression level was detected by Western blot，the proliferation ability by MTT，the secretion ability after glucose stimulation by ELISA. Results：The pawr genes were verified by gene sequencing. The recombinant plasmid was successfully extracted by high purity plasmid extraction median kit. The recombinant adenovirus vector plasmid with mono-restriction endonuclease enzyme was evaluated. The earliest expression of GFP was observed in HEK293 cells and after 48 h of amplification the CPE was observed and the final viral titer was 1×109 pfu/ml. The protein expression level of PAR-4 was significantly increased in low glucose+overexpression of PAR-4 group and high glucose+overexpression of PAR-4 group than in its control groups. Proliferation ability and insulin secretion were decreased in high glucose+overexpression of PAR-4 group than in low glucose+overexpression of PAR-4 group and high glucose control group. Conclusion：The over-expres－sion of PAR-4 group has higher expression of PAR-4. After transfection and culture by high glucose concentration，the pro－liferation ability of high-glucose cultured NIT-1 is decreased.
