Expression of the human interleukin-2 receptor gamma chain in insect cells using a baculovirus expression vector.
The gene encoding the gamma-chain of the human 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 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 gamma-receptor specific antiserum. Metabolic labelling with [35S]-methionine and SDS-PAGE analysis of the recombinant baculovirus infected insect cells verified the immunoblotting data. The expressed IL-2R gamma- protein could also be determined on the surface of infected insect cells by flow cytometer analysis.
Raivio
E
E
Abo Akademi University, Department of Biochemistry and Pharmacy, Finland.
Oetken
C
C
Oker-Blom
C
C
Engberg
C
C
Akerman
K
K
Lindqvist
C
C
D000818:Animals; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D008970:Molecular Weight; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D018411:Spodoptera
D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins