Authentication And Bioinformatics Analysis About A Possible Downstream Gene Of P53 Protein-PAP1

The p53 gene is the most correlative suppress cancer gene with human's tumor.The past research reveal that p53 gene act by the P53 protein being combined with the p53 consensus binding sequence in genes, activating the downstream gene to be translated,act. Authenticate the P53 downstream genes is the key to understands adjustor and control network of p53 gene.In past work,we the establish a wild-type p53 gene cells——U251-pTet-p53.Under the control of tetracycline-response element,induce p53 gene to express.we get a new possible p53 downstream gene-PAP1 gene by the difference revealment technique.In this paper,We construct pET28b-p53 recombinant expression plasmid according to the designed technology track, and transform to E.coli BL21 (DE3), induced with IPTG.. PAP1 gene is marked by Digoxin as a probe,then electrophoretic mobility shift assay is experiment between P53 protein and consensus binding sequence clone from PAP1 gene to authenticate PAP1.We also do gene structure analysis and function analysis to the PAP1 gene with bioinformatics method.The result of our study reveal : The protein encoded by PAP1 gene belong to a member of the immunoglobulin superfamily(IGSF). Similarity Search of GenBank database shows that the high similarity bewteen PAP1mRNA and DORA/DREV1 mRNA. Then, pairs alignment between PAP1 and DREV1 finds that their similarity is 80.8%. DREV1 protein belong to a methyl transferase by Interpro server search for protein family class . DREV1 protein includes a domain of methyl transferase and PAP1 protein includes it. So we guess that they are the same gene . Alignment of the predicted protein sequence for Human, Pan troglodytes, Canis, Mus musculus and Gallus gallus revealed it was highly conserved. Appearance of the bioinformatics,we can reveal some rule in a large of biological data and deal with the data .