| Vaccinia Virus (VV) belongs the to the Poxviridae family of viruses. It is a large, double-stranded DNA virus that encodes more than 200 viral proteins. The virus completes its replication cycle in the cytoplasm of the host cell, never entering the nucleus. Gene expression in the virus is temporally regulated, with the production of early, intermediate and late proteins.;One of the open reading frames of VV, designated F18R in the Western Reserve strain of the virus, encodes an essential, small, highly basic 11-kDa protein known as VP11. The protein is expressed after viral replication, thereby making it a late protein. There are ten Serine residues on VP11, two of which are thought to be phosphorylated. These sites of phosphorylation lie within PSSP motifs---sequences of amino-acids that serve as recognition sites for certain proline-directed kinases. The primary structure of VP11 highlights another interesting aspect of its nature---its characterization as possibly an intrinsically disordered protein.;Through the use of various bioinformatic tools, the disordered nature of VP11 is probed, in addition to substantiation of the possible sites of phosphorylation of the protein. A corroboration of disordered regions and phosphorylation sites was also examined. Additionally, differences in bacterially expressed VP11 versus natively expressed VP11 were investigated, with specific regard to the behavior of phosphorylated VP11 on certain chromatographic media Finally, in an attempt to produce substantial amounts of full-length soluble protein, VP11 was expressed bacterially using temperatures lower than those typical of normal bacterial culture. |
