Expression of recombinant human coagulation factor VII by the Lizard Leishmania expression system.

Sina Mirzaahmadi (1) , Golnaz Asaadi-Tehrani (1) , Mojgan Bandehpour (2) , Nooshin Davoudi (3) , Leila Tahmasbi (4) , Nahid Hosseinzadeh (2) , Hasan Mirzahoseini (3) , Kazem Parivar (1) , Bahram Kazemi (2, 5)

1 Department of Biology, Science and Research Branch
2 SBU - Cellular and Molecular Biology Research Center
3 Institut Pasteur d'Iran
4 Department of Hematology
5 Biotechnology Department

The variety of recombinant protein expression systems have been developed as a resource of FVII gene expression. In the current study, the authors used a novel protein expression system based on the Iranian Lizard Leishmania, a trypanosomatid protozoan as a host for expression of FVII. Plasmid containing cDNA encoding full-length human FVII was introduced into Lizard Leishmania and positive transfectants were analyzed by SDS-PAGE and Western blot analysis. Furthermore, biological activity of purified protein was detected by PT assay. The recombinant strain harboring a construct was analyzed for expression of FVII at the mRNA and protein level. Purified rFVII was obtained and in order to confirm the purified compound was in fact rFVII. Western blot analysis was carried out. Clotting time in PT assay was reduced about 30 seconds with the purified rFVII. In Conclusion, this study has demonstrated, for the first time, that Leishmania cells can be used as an expression system for producing recombinant FVII.

Mots clés: recombinant; human coagulation factor VII; Lizard Leishmania; expression system

Domaines: Biotechnologie; Biotechnologies

Sina Mirzaahmadi, Golnaz Asaadi-Tehrani, Mojgan Bandehpour, Nooshin Davoudi, Leila Tahmasbi, et al.. Expression of recombinant human coagulation factor VII by the Lizard Leishmania expression system.. Journal of Biomedicine and Biotechnology, 2011, 2011, pp.873874. ⟨10.1155/2011/873874⟩. ⟨pasteur-00750784⟩