关键词: 腺病毒, 耐辐射球菌, PprI基因

Abstract: PprI gene has obvious prevention and control effect on acute radiation damage caused by neutron and gamma rays in mammals. In this study, the PprI gene was directed cloned into the shuttle plasmid pADV-mCMV-MCS-3xFLAG using the non-replicating adenovirus vector system AdMax packaging system, and then co-transfected into HEK293 cells with the adenovirus skeleton plasmid for homologous recombination. The recombinant adenovirus pAdMax-PprI was obtained and identified. The PprI protein was bioinformatics analyzed. PCR results of recombinant adenovirus pAdMax-PprI showed that the inserted fragment was consistent with the size of the PprI gene fragment, and sequencing showed that the inserted fragment was consistent with the PprI gene sequence. Western blot results showed that PprI protein was immunogenic with a molecular weight of about 45 kDa. Bioinformatics analysis showed that PprI gene encodes a hydrophilic protein composed of 163 amino acid residues, which exists in cell membrane, has no signal peptide and transmembrane structure, and has 5 antigenic epitopes. The secondary structure includes three forms of random curling, α-helix and β-folding. Random curl (46.73%) and α helix (28.83%) were the main structures. The prediction results of tertiary structure were consistent with those of secondary structure. The results laid a foundation for further studies on the function of PprI and its radiation resistance.

Key words: adenovirus, radiococcus durans, PprI gene