Document Detail


Expression of the mouse interleukin-2 receptor gamma chain in insect cells using a baculovirus expression vector--comparison with the human common gamma chain.
MedLine Citation:
PMID:  9042425     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The gene encoding the gamma-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-2R gamma. Kinetic analysis demonstrated that the corresponding protein could be detected as an approximately 50 kDa protein already at 24 h post-infection. Intrinsic labelling with [35S]-methionine/cysteine 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 cytometric analysis. Comparison of the molecular weights between baculovirus expressed human and mouse IL-2R gamma chains indicated differences in the glycosylation pattern despite similar numbers of N-linked glycosylation sites.
Authors:
K Stenroos; A West; E Raivio; C Lindqvist
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Scandinavian journal of immunology     Volume:  45     ISSN:  0300-9475     ISO Abbreviation:  Scand. J. Immunol.     Publication Date:  1997 Feb 
Date Detail:
Created Date:  1997-03-26     Completed Date:  1997-03-26     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0323767     Medline TA:  Scand J Immunol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  140-4     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Abo Akademi University, Turku, Finland.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cells, Cultured
Electrophoresis, Polyacrylamide Gel
Flow Cytometry
Gene Expression Regulation*
Genetic Vectors / genetics
Glycosylation
Humans
Immunoblotting
Lepidoptera
Mice
Nucleopolyhedrovirus / genetics
Promoter Regions, Genetic
Receptors, Interleukin-2 / genetics*
Recombinant Proteins / genetics,  immunology
Recombination, Genetic
Viral Proteins / genetics
Viral Structural Proteins
Chemical
Reg. No./Substance:
0/Receptors, Interleukin-2; 0/Recombinant Proteins; 0/Viral Proteins; 0/Viral Structural Proteins; 0/polyhedrin protein, Nucleopolyhedrovirus

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