Expression in bacteria of a polypeptide encoded by a transforming fragment of herpes simplex virus type 2.
We have constructed a plasmid (pMD2) containing the 38,000 MW polypeptide (38K polypeptide) gene from the transforming Bg1II-N fragment of HSV-2 fused to the amino-terminal portion of the beta-galactosidase gene in plasmid pUC8. Nucleotide sequence determination around the fusion-junction confirmed that the viral gene sequences starting at 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 against the 38K viral polypeptide and antibodies raised against a synthetic peptide corresponding to the nine C-terminal amino acid residues of the 38K viral protein, detected the fusion protein in bacteria containing the recombinant plasmid pMD2 but not in Escherichia coli containing a related plasmid or no plasmid.
Danik
M
M
Centre de Recherche, Hôpital Maisonneuve-Rosemont, Montreal, Quebec, Canada.
Suh
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D000818:Animals; D001483:Base Sequence; D006224:Cricetinae; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D005814:Genes, Viral; D006561:Herpes Simplex / Q000235:genetics*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D014162:Transfection; D014170:Transformation, Genetic / Q000502:physiology; D014760:Viral Fusion Proteins / Q000235:genetics*
D014760:Viral Fusion Proteins