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Expression of the Mouse Interleukin‐2 Receptor Gamma Chain in Insect Cells Using a Baculovirus Expression Vector—Comparison with the Human Common Gamma Chain
Author(s) -
STENROOS K.,
WEST A.,
RAIVIO E.,
LINDQVIST C.
Publication year - 1997
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1046/j.1365-3083.1997.d01-378.x
Subject(s) - polyhedrin , sf9 , spodoptera , biology , recombinant dna , microbiology and biotechnology , baculoviridae , autographa californica , nuclear polyhedrosis virus , recombinant virus , virology , gene , virus , biochemistry
The gene encoding the γ‐chain of the mouse Interleukin‐2 receptor was expressed in lepidopteran insect cells using the baculovirus expression vector system. The corresponding gene was inserted under the polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus and expressed in the Spodoptera frugiperda insect cell line Sf9 during viral infection. The recombinant receptor protein was identified by immunoblotting in cell lysates prepared from insect cells infected with the produced recombinant virus VL1392‐mIL‐2Rγ. Kinetic analysis demonstrated that the corresponding protein could be detected as an ≈50 kDa protein already at 24 h post‐infection. Intrinsic labelling with [ 35 S]‐methionine/cysteine and SDS‐PAGE analysis of the recombinant baculovirus infected insect cells verified the immunoblotting data. The expressed IL‐2Rγ protein could also be determined on the surface of infected insect cells by flow cytometric analysis. Comparison of the molecular weights between baculovirus expressed human and mouse IL‐2Rγ chains indicated differences in the glycosylation pattern despite similar numbers of N‐linked glycosylation sites.