Incorporation of chimeric Gag protein into retroviral particles

R. A. Weldon, C. R. Erdie, M. G. Oliver, J. W. Wills

Research output: Contribution to journalArticlepeer-review

87 Scopus citations

Abstract

The product of the Rous sarcoma virus (RSV) gag gene, Pr76(gag), is a polyprotein precursor which is cleaved by the viral protease to yield the major structural proteins of the virion during particle assembly in avian host cells. We have recently shown that myristylated forms of the RSV Gag protein can induce particle formation with very high efficiency when expressed in mammalian cells (J.W. Wills, R.C. Craven, and J.A. Achacoso, J. Virol. 63: 4331-4343, 1989). We made use of this mammalian system to examine the abilities of foreign antigens to be incorporated into particles when fused directly to the myristylated Gag protein. Our initial experiments showed that removal of various portions of the viral protease located at the carboxy terminus of the RSV Gag protein did not disrupt particle formation. We therefore chose this region for coupling of iso-1-cytochrome c from Saccharomyces cerevisiae to Gag. his was accomplished by constructing an in-frame fusion of the CYC1 and gag coding sequences at a common restriction endonuclease site. Expression of the chimeric gene resulted in synthesis of the Gag-cytochrome fusion protein and its release into the cell culture medium. The chimeric particles were readily purified by simple centrifugation, and transmission electron microscopy of cells that produced them revealed a morphology similar to that of immature type C retrovirions.

Original languageEnglish (US)
Pages (from-to)4169-4179
Number of pages11
JournalJournal of virology
Volume64
Issue number9
DOIs
StatePublished - 1990

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Fingerprint Dive into the research topics of 'Incorporation of chimeric Gag protein into retroviral particles'. Together they form a unique fingerprint.

Cite this