EXPRESSION OF THE MOUSE INTERLEUKIN-2 RECEPTOR-GAMMA CHAIN IN INSECT CELLS USING A BACULOVIRUS EXPRESSION VECTOR - COMPARISON WITH THE HUMAN COMMON GAMMA-CHAIN
K. Stenroos et al., EXPRESSION OF THE MOUSE INTERLEUKIN-2 RECEPTOR-GAMMA CHAIN IN INSECT CELLS USING A BACULOVIRUS EXPRESSION VECTOR - COMPARISON WITH THE HUMAN COMMON GAMMA-CHAIN, Scandinavian journal of immunology, 45(2), 1997, pp. 140-144
The gene encoding the gamma-chain of the mouse Interleukin-2 receptor
was expressed in lepidopteran insect cells using the baculovirus expre
ssion vector system. The corresponding gene was inserted under the pol
yhedrin promoter of the Autrographa californica nuclear polyhedrosis v
irus and expressed in the Spodoptera frugiperda insect cell line Sf9 d
uring 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 anal
ysis demonstrated that the corresponding protein could be detected as
an approximate to 50 kDa protein already at 24 h post-infection, Intri
nsic labelling with [S-35]-methionine/cysteine and SDS-PAGE analysis o
f the recombinant baculovirus infected insect cells verified the immun
oblotting data, The expressed IL-2R gamma protein could also be determ
ined on the surface of infected insect cells by flow cytometric analys
is. Comparison of the molecular weights between baculovirus expressed
human and mouse IL-2R gamma chains indicated differences in the glycos
ylation pattern despite similar numbers of N-linked glycosylation site
s.