Synthesis of the outer-capsid glycoprotein of the simian rotavirus SA11 in Escherichia coli.

The major outer layer protein, VP7, of the simian rotavirus SA11 has been synthesized in Escherichia coli, under the control of the lac promoter, as a fusion polypeptide with beta-galactosidase (beta Gal). The viral protein in the hybrid polypeptide is missing its N-terminal hydrophobic region and 26 amino acids (aa) at its C-terminus; it is flanked at both ends by beta Gal sequences. We have purified the hybrid 145-kDa protein by affinity chromatography using a column specific for beta Gal. Unexpectedly, a second protein of 118-kDa was also specifically bound to the column. N-terminal aa sequence analysis of these two proteins showed that the 145-kDa protein represented the expected fusion product, whereas the 118-kDa protein was apparently the result of initiation of translation at an internal site close to the 3' end of the viral sequence, in the chimeric mRNA. Each of the two polypeptides represented about 2 to 3% of the total protein of the recombinant-plasmid-carrying bacteria. When a bacterial lysate enriched for the hybrid polypeptides was injected into mice, it induced neutralizing antibodies to SA11 rotavirus.