Vietnam Academy of Science and Technology

Project's information

Project's title	A mammalian cell culture system for expression of clotting factors
Research hosting institution	Institute of Biotechnology
Project leader’s name	Dr. Nong Van Hai
Project duration	01/01/2009 - 01/01/2010
Project’s budget	300 000 000 VND
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Goal and objectives of the project	Cloning cDNAs encoding human coagulation factors F8 and F9; expression of recombinant F9 in mammalian cells (CHO) and determination of recombinant F9 activity.
Main results	*Scientific aspects* Collection of human liver tissue Extraction of total RNA Cloning and sequencing F8, F9 Vector construction for expression of the F9 Expression of F9 in mammalian cells (CHO) Condition optimization for culturing CHO cells Transfection of recombinant vector into CHO Cloning stable CHO cells containing the F9 gene Purification of recombinant F9 Confirmation of F9 using Western blot Determination of F9 activity
Novelty and actuality and scientific meaningfulness of the results	Cloning genes encoding coagulation factors F8/F9 and F9 expression have not been carried out in Vietnam yet. Therefore, the scientific result of the project will be the foundation for the expression of functional protein F8 and production of recombinant protein F9, which has medical applications. The outcomes of the project help direct other relating researches and initiate the application of molecular techniques to pharmaceutical drugs.
Products of the project	Scientific papers in referred journals (listed) Nguyen Thuy Duong, Nguyen Van Phong, Nong Van Hai (2010). Cloning and sequence analysis of gene encoding human coagulation factor IX. Journal of Biotechnology 8 (4):1785-1792 Nguyen Thuy Duong, Nguyen Van Phong, Nong Van Hai (2011). Vector construction for expression of the human factor IX. Journal of Biotechnology 9 (3): 289-295 Specific products 12 recombinant plasmids containing 6 cDNA fragments encoding 6 domains of F8: A1, A2, B, A3, C1 và C2 Three recombinant plasmids containing full-length cDNA encoding F9 One recombinant expression plasmid containing full-length cDNA encoding F9 Obtaining recombinant protein F9 expected to be close to the biologically active Other products (If applicable) Submission of three cDNA sequences encoding F9 to GenBank with accession numbers: FR846238, FR846239 và FR846240 Condition optimization for culturing CHO
Recommendations	The study and development of recombinant protein F8/F9 has scientific value and potential for future application. Because this study has only created F8 and F9 gene sources and designed preliminary study of the expression of F9. We propose further perfecting studies of the expression of F9.