Expression in bacteria of a polypeptide encoded by a transforming fragment of herpes simplex virus type 2
Danik, M.; Suh, M.
Cancer Detection and Prevention 15(2): 107-113
1991
ISSN/ISBN: 0361-090X PMID: 2032251 Document Number: 376886
We have constructed a plasmid (pMD2) containing the 38,000 MW polypeptide (38K polypeptide) gene from the transforming BglII-N fragment of HSV-2 fused to the amino-terminal portion of the .beta.-galactosidase gene is plasmid pUC8. Nucleotide sequence determination around the fusion-junction confirmed that the viral gen sequences starting as its second codon is in the correct reading frame in relation to the translation initiation codon of .beta.-galactosidase. The lac control sequences direct the synthesis of a 39K protein. This protein was shown to be structurally related to the 38K protein from HSV-2-infected cells by partial proteolytic cleavage analysis. Furthermore, antiserum directed against HSV-2-infected cells, as well as a monoclonal antibody agains the 38K viral polypepide and antibodies raised againist a synthetic peptide corresponding to the nine C-terminal amino acid residues of the 38K virual protein, detected the fusion protein in bacteria containing the recombinant plasmid pMD2 but not in Escherichia coli containing a related plasmid or no plasmid.