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Virologica Sinica, 17 (4) : 340-343, 2002
Research Article
Expression,Purification and Bioactivity Detection of Recombinant Hum an IL-12 in Argyogramma agnata
Institute of Virology.Wuhan University。Wuhan 430072。China
 Correspondence:
(155.77KB)  
Abstract
Two cDNA fragments encoding human interleukin一12(hII一12)P35 and P40 subunits were
isolated by RT.PCR from KB cells stimulated with PDBu,and then cloned into pCR .1 respcetively
and down the double promoters of pAcUw 5 1.The recombinant baculovirus Ac—hiL2 was obtained by
cotransfecting with pAeUW 5 1一ILl2 and baeuloGold M Linearized baculovirus DNA.Argyrogramma
agnata larvae were infected with recombinant baculovirus Ac—hiLl2 by hemoeoel injection.The rhII,
1 2 was purified from the supernatant with affinity chromatography.The blood lymph supernatant har—
vested and rhII,12 purified were SUbjected to SDS-PAGE(silver stain)and Western blot.The level of
rhIL-12 was detected by EU SA.Bioactivity of purified rhII,12 samples were detected by M 1vr
method.It is indicated that Ae-hlL12 can replicate in fat body and midgut cells of Argyrogramma
Agnata larvae.The MW of rhlL一12 expressed was 75kD.The expresion levels of rhIL-12 were 17.8
/~g/lO cells and 200—300mg/L in Sf9 cells and Argyrogramma agnata Staudinger larvae blod
lymph respectively.The purified rhII,12 has significant activities which enhanced NK eytotoxieity
and increased the proliferation of human PBMC PHA-P activated significantly.
  Published online: 5 Nov 2002
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