Expression of bioactive human interferon-gamma in transgenic rice cell suspension cultures.
We investigated the possibility of producing the therapeutic recombinant cytokine, Interferon-gamma (IFN-gamma), in transgenic rice cell (Oryza sativa, cultivar TNG67) suspension cultures. We tested expression of two vector constructs, each harboring an alphaAmy3 leader peptide and a C-terminus His 6 tag fused to a human IFN-gamma cDNA, one driven by a sucrose-starvation inducible promoter (rice alphaAmy3 promoter) and the other by a constitutive maize ubiquitin promoter, in rice cell suspensions, introduced via Agrobacterium tumefaciens. There was a significant difference in the amounts of recombinant IFN-gamma protein produced by the Ups and Amy cell lines, as cytosolic and secretory proteins respectively. Immunological analysis of IFN-gamma recombinant protein conferred a dose-dependent anti-dengue virus activity in human A549 cells, similar to the commercial product. We discuss the attractive attributes of using rice cell suspension system for the expression of therapeutic recombinant IFN-gamma.
Chen
Tzy-Li
TL
Institute of BioAgricultural Sciences, Academia Sinica, Taipei 115, Taiwan, Republic of China.
Lin
Yi-Ling
YL
Lee
Yi-Ling
YL
Yang
Ning-Sun
NS
Chan
Ming-Tsair
MT
D018929:Cell Culture Techniques; D003716:Dengue Virus / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D012275:Oryza / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins; D012231:Rhizobium / Q000235:genetics; D014407:Tumor Cells, Cultured; D025801:Ubiquitin / Q000235:genetics
D011994:Recombinant Proteins; D025801:Ubiquitin; D007371:Interferon-gamma