title: [Eukaryotic expression of recombinant porcine single-chain IL-12 and its biologic  activity].
abstract: OBJECTIVE
  To express the recombinant porcine single-chain interleukin-12 (pscIL-12) gene in CHO-K1 cells,  and identify biological activity of pscIL-12 fusion protein.
  
  
  METHODS
  The recombinant pcDNA3.1(+)-pscIL-12 plasmid was transfected into the CHO-K1 cells using Sofast(TM);  reagent. The levels of pscIL-12 fusion protein and IFN-γ produced by human peripheral blood mononuclear cells (PBMCs) stimulated with the supernatant of CHO-K1 cells were detected by ELISA. NK cell cytotoxicity was tested by MTT assay. The lymphocyte proliferation was examined by flow cytometry.
  
  
  RESULTS
  ELISA showed that the transfected CHO-K1 cells had a relatively higher expression of pscIL-12 fusion protein. And the pscIL-12 fusion protein possessed strong bioactivities in enhancing the NK cell cytotoxicity,  increasing the IFN-γ production,  and stimulating the lymphocyte proliferation.
  
  
  CONCLUSIONS
  CHO-K1 cells transfected with pcDNA3.1(+)-pscIL-12 recombinant plasmid can successfully express pscIL-12 fusion protein,  which has the expected biological activities.
authors: Department of Microbiology and Parasitology, Bengbu Medical College, China. wangxuenm0508@sina.com
authors: Xue-mei
authors: XM
authors: Wang
authors: Yuan-ying
authors: YY
authors: Yuan
authors: Qiang
authors: Q
authors: Fang
authors: Hui
authors: H
authors: Xia
authors: Xin
authors: X
authors: Sun
authors: Bai-qing
authors: BQ
authors: Li
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D015870:Gene Expression; D007371:Interferon-gamma / Q000096:biosynthesis; D018664:Interleukin-12 / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000187:drug effects / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013552:Swine
chemical_list: D011993:Recombinant Fusion Proteins; D018664:Interleukin-12; D007371:Interferon-gamma