The Research Of Interaction Between Of The Human Melanoma Associated Antigen-b18and Human Nf-κb Inhibitors Tnip1

Objective:To observe the interaction of Human melanoma associated antigen-B18between the Human NF-κB inhibitors TNIPl,and indentify the binding site of the MAGE-B18and TNIP1.Reaearch the sub-signal transduction way of them and prepared experimental ground for further study of the two protein.Method:Transfect the293T with the eukaryotic expressing vector of MAGE-B18and TNIP1.Using the co-immunoprecipitation to verify the interaction between the two protein. After cofirming the interaction, using the PCR method to construct the mutation of the two protein with HA or Myc tag and expressing in the293T cell. Using co-IP to identify the binding site of the two protein. Using western-blotting to observe the sub-signal transduction. Using FCM to observe the apoptosis rate of293T cell.Result:The result of co-IP show that the TNIP1and MAGE-B18protein can be binding each other in the cell. Successfully construction the mutation of TNIPl and MAGE-B18with Tag, and expression in the293T cell. Indentify the binding site of TNIP1is311-535aa, and MAGE-B18is114-284aa. The result of western-blotting show that the TNIP1and MAGE-B18protein can be co-working to influence the NF-kBThe result of FCM show that the TNIP1and MAGE-B18protein can be co-working to inhibite the cell apoptosis.Conclusion:The human melanoma associated antigen B18can be interaction with human NF-κB inhibitors TNIP1in the cell. The TNIP1can be co-working with MAGE-B18to inhibit the NF-κB. At the same time, the MAGE-B18can be co-working with the TNIP1to inhibit the cell apoptosis. This research prepared experimental ground for further study of the MAGE-B18and TNIP1in the tumor formation and progress.