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Expression of the Human Interleukin–2 Receptor Gamma Chain in Insect Cells Using a Baculovirus Expression Vector
Author(s) -
RAIVIO E.,
OETKEN C.,
OKERBLOM C.,
ENGBERG C.,
ÅKERMAN K.,
LINDQVIST C.
Publication year - 1995
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 - 10.1111/j.1365-3083.1995.tb03576.x
Subject(s) - vector (molecular biology) , insect , biology , expression (computer science) , microbiology and biotechnology , receptor , virology , genetics , gene , recombinant dna , botany , computer science , programming language
The gene encoding the γ‐chain of the human lnterleukin–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 recombinant virus. At 40 h post infection the corresponding protein was detected as two major bands with apparent molecular weights of 50–60 kDa using a rabbit anti–human IL–2R γ‐receptor specific antiserum. Metabolic labelling with [ 35 S]–methionine 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 cytometer analysis.