| 研究报告 |
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| Research on the Expression of Human Coagulation Factor IX in Tobacco |
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Abstract Hemophilia B is a bleeding disorder caused by deficiency or abnormality in the plasma coagulation protein factor IX. Currently treatment method of hemophilia B is based on protein replacement therapy by blood transfusion and intravenous infusion of condensate of coagulation protein factor IX with excellent therapy effect, which also has some problems including expensive costs, and high risks of virus infection by blood filtering. So how to obtain human factor IX more cheaply and safely is very important for the cure of hemophilia B. Plant expression system own advantage on cost and security, which showed good potential for production of human factor IX. Therefore, in this work, plant binary expression vector p35S-2300：：hFIX（2.8kb）：：noster was constructed and introduced into Agrobacterium tumefaciens strain EHA105 by freezing-melting method. The resulting vector was introduced into tobacco（Nicotiana tabacum L. cv. Bai Rihong）plants via Agrobacterium tumefaciens -mediated method. Total 4 independent transgenic tobacco plants confirmed by PCR and Southern blot analysis were obtained. The hFIX gene was introduced into genome of transgenic tobacco plants, ranging from 1 to 4 copies. The expression of the introduced gene in transgenic T0 plants was confirmed by RT-PCR and ELISA analysis at transcript and translation level, respectively. Content of hFIX ranged from 2.5 to 8.8ng/g·FW in fresh leaves of transgenic tobacco with immunocompetence. This work provides good foundation for further expression of hFIX and is also helpful for expression of other pharmaceutical proteins using plant expression system.
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Received: 10 February 2006
Published: 25 March 2006
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