The characterisation of a cervine immunoregulatory cytokine, interleukin 12.
The cloning, sequencing, and production of cervine interleukin-12 is described. The cervine IL-12 p35 subunit coding sequence is 666 bp long and has highest homology to bovine p35 (94%), followed by human (79%), then murine (57%). The cDNA codes for a 221 aa long protein with predicted molecular weight of 24,902 Da. The cervine p40 subunit has a coding sequence of 984 bp and shows 96% homology to bovine, 85% homology to human, and 65% homology to murine p40 respectively. Cervine p40 cDNA codes for a 327 aa long protein with a predicted molecular weight of 37,461. Both subunits were inserted into a recombinant baculovirus that was then used to produce cervine IL-12 in Trichoplusia ni cells. Interleukin-12 was secreted into the culture medium and was biologically active as measured by proliferation of mitogen sensitised peripheral blood lymphocytes and the induction of interferon-gamma transcription in peripheral blood lymphocytes.
Lockhart
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E
Department of Microbiology, University of Otago, Dunedin, New Zealand.
Slobbe
L
L
Buchan
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D018076:DNA, Complementary; D003670:Deer / Q000235:genetics*; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics; D018664:Interleukin-12 / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D018664:Interleukin-12; D007371:Interferon-gamma