title: Expression of human coagulation Factor IX in transgenic tomato (Lycopersicon esculentum).
abstract: In the present study, a plant binary expression vector PG-pRD12-hFIX (where PG is polygalacturonase) harbouring the hFIX (human coagulation Factor IX) gene was constructed and introduced into tomato (Lycopersicon esculentum) via Agrobacterium tumefaciens-mediated transformation. After kanamycin selection, 32 putative independent transgenic tomato plants were regenerated. PCR and Southern-blot analyses confirmed the transgenic status of some plants. RT (reverse transcription)-PCR analysis for the expression of the introduced gene (hFIX) demonstrated that the hFIX gene was expressed specifically in fruits of the tomato. Western-blot analysis confirmed the presence of a 56 kDa band specific to hFIX in the transformed tomatoes. ELISA results showed that the expression of hFIX protein reached a maximum of 15.84 ng/g fresh weight in mature fruit. A blood-clotting assay demonstrated the clotting activity of the expressed hFIX protein in transgenic tomato fruits. This is the first report on the expression of hFIX in plants, and our research provides potentially valuable knowledge for further development of the plant-derived therapeutic proteins.
authors.affiliation: Plant Biotechnology Research Center, School of Agriculture and Biology, Fudan-Shanghai Jiaotong University (SJTU)-Nottingham Plant Biotechnology R&D Center, SJTU, Shanghai 200030, People's Republic of China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Lingxia
authors.initials: L
authors.lastname: Zhao
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Chen
authors.forename: Lijie
authors.initials: L
authors.lastname: Cui
authors.forename: Weiwei
authors.initials: W
authors.lastname: Ren
authors.forename: Kexuan
authors.initials: K
authors.lastname: Tang
mesh_terms: D005164:Factor IX / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018551:Solanum lycopersicum / Q000502:physiology*; D030821:Plants, Genetically Modified / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D005164:Factor IX