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

Carlos F. Arias, Teresa Ballado, Magda Plebafiski

Research output: Contribution to journalArticleResearchpeer-review

19 Citations (Scopus)

Abstract

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 β-galactosidase (βGal). 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 βGal sequences. We have purified the hybrid 145-kDa protein by affinity chromatography using a column specific for βGal. 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.

Original languageEnglish
Pages (from-to)211-219
Number of pages9
JournalGene
Volume47
Issue number2-3
DOIs
Publication statusPublished - 1 Jan 1986

Keywords

  • lac promoter
  • neutralizing antibodies
  • Recombinant DNA
  • subunit vaccine
  • VP7 glycoprotein
  • β-galactosidase fusion

Cite this

Arias, Carlos F. ; Ballado, Teresa ; Plebafiski, Magda. / Synthesis of the outer-capsid glycoprotein of the simian rotavirus SA11 in Escherichia coli. In: Gene. 1986 ; Vol. 47, No. 2-3. pp. 211-219.
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abstract = "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 β-galactosidase (βGal). 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 βGal sequences. We have purified the hybrid 145-kDa protein by affinity chromatography using a column specific for βGal. 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.",
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Synthesis of the outer-capsid glycoprotein of the simian rotavirus SA11 in Escherichia coli. / Arias, Carlos F.; Ballado, Teresa; Plebafiski, Magda.

In: Gene, Vol. 47, No. 2-3, 01.01.1986, p. 211-219.

Research output: Contribution to journalArticleResearchpeer-review

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