Expression of human coagulation Factor IX in transgenic tomato (Lycopersicon esculentum).
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.
Zhang
Hui
H
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.
Zhao
Lingxia
L
Chen
Yuhui
Y
Cui
Lijie
L
Ren
Weiwei
W
Tang
Kexuan
K
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
D011994:Recombinant Proteins; D005164:Factor IX