Murine interleukin-4 production with baculovirus: an easy and rapid method for a small scale production of functional interleukins.
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.
Cottrez
F
F
Unite CNRS-URA 1854, Institut Pasteur de Lille, France.
Auriault
C
C
Capron
A
A
Kusznier
J P
JP
Groux
H
H
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005822:Genetic Vectors; D006031:Glycosylation; D015847:Interleukin-4 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D018411:Spodoptera
D018076:DNA, Complementary; D011994:Recombinant Proteins; D015847:Interleukin-4