Murine interleukin-4 production with baculovirus: an easy and rapid method for a small scale production of functional interleukins

Eur Cytokine Netw. 1994 Sep-Oct;5(5):481-7.

Abstract

We described a baculovirus expression system for high level production of secreted murine recombinant IL-4. We have constructed a recombinant baculovirus based on Autographa californica polyhedrosis virus, containing both a synthetic PCR-derived murine IL-4 cDNA under the control of the polyhedrin promoter and the lacZ gene under the control of the P10 promoter to allow an easy detection of recombinant virus. The baculovirus IL-4 was fully functional in biological assay and was present under two glycosylated forms in the supernatants of infected Sf9 cells. We also detected a third unglycosylated intracytoplasmic form resulting from a fusion between the 35 first amino acids of polyhedrin and the murine IL-4. Finally, confocal microscopy showed that this recombinant protein was secreted along a classical pathway like in mammalian cells.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Base Sequence
  • Cell Line
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Fluorescent Antibody Technique
  • Gene Expression
  • Genetic Vectors
  • Glycosylation
  • Interleukin-4 / biosynthesis*
  • Interleukin-4 / chemistry
  • Interleukin-4 / genetics*
  • Mice
  • Molecular Sequence Data
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Saccharomyces cerevisiae / genetics
  • Spodoptera

Substances

  • DNA, Complementary
  • Recombinant Proteins
  • Interleukin-4