[Expression of human interleukin-12 in Baculovirus expression system].
Human Interleukin-12 (hIL-12) is a key immunoregulatory cytokine and the only heterodimeric cytokine found by far. It is a disulfide-linked heterodimer composed of a 35 kD light chain (P35) and a 40 kD heavy chain (P40). Two recombinant transfer vector plasmids pAcAB3-p35 (including hIL-12 p35 gene) and pAcAB3-p40 (including hIL-12 p40 gene) were constructed and co-transfected the insect cells Sf9 with AcNPV BaculoGold Linearized Baculovirus DNA. The obtained recombinant viruses, AcNPV-OCC(-)-hiL-12 (p35) and AcNPV-OCC(-)-hIL-12 (p40), were used to infect the Sf9 cells separately, and the supernatants of culture and cell lysate were examined by using SDS-PAGE and Western blot. Results showed that both hIL-12 p35 and p40 gene were successfully expressed in insect cells and secreted. The expression productions had immunogenicity.
Yang
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Biopharmeutical Center, State Key Lab for Biocontrol, Zhongshan University, Guangzhou 510275, China.
Zhang
X
X
Long
X
X
Chen
X
X
Wang
X
X
Li
Y
Y
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D018664:Interleukin-12 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics; D018411:Spodoptera / Q000166:cytology / Q000378:metabolism*; D014162:Transfection
D004274:DNA, Recombinant; C508858:IL12A protein, human; D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D021122:Protein Subunits; D018664:Interleukin-12