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Expression of human coagulation Factor IX in transgenic tomato ( Lycopersicon esculentum )
Author(s) -
Zhang Hui,
Zhao Lingxia,
Chen Yuhui,
Cui Lijie,
Ren Weiwei,
Tang Kexuan
Publication year - 2007
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20060224
Subject(s) - lycopersicon , agrobacterium tumefaciens , genetically modified crops , biology , transgene , genetically modified tomato , western blot , microbiology and biotechnology , expression vector , recombinant dna , factor ix , gene , botany , biochemistry
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.