Expression and purification of myxoma virus leukemia associated protein N-terminal domain

CHED 840

Rebecca Asiamah, zynbiec@gmail.com and Dana C. Lawrence, lawrence@hood.edu. Department of Chemistry and Physics, Hood College, 401 Rosemont Ave, Frederick, MD 21701
Myxoma virus Leukemia Associated Protein (MV-LAP) is a protein encoded by Myxoma virus, a poxvirus responsible for the lethal disease, myxomatosis in the European Rabbit. MV-LAP has developed a “stealth” mechanism to evade the host immune system by reducing the expression of major histocompatibility complex (MHC)-1 molecules, responsible for identifying self vs. non-self antigens. In order to help understand this stealth mechanism, the N-terminal domain (NTD; 94 residues) of MV-LAP will be characterized structurally using NMR. The current project involves the cloning, expression and purification of this protein. The MV-LAP NTD was cloned as two different fusion proteins, covalently attached to Maltose Binding Protein and NusA. Multiple growth temperatures were screened, and proteolytic cleavage and subsequent purification of the MV-LAP NTD was successful; preparation of a sample for NMR analysis is currently underway.