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Expression of Recombinant Human Coagulation Factor VII by the LizardLeishmaniaExpression System

Authors :
Kazem Parivar
Sina Mirzaahmadi
Nahid Hosseinzadeh
Bahram Kazemi
Hasan Mirzahoseini
Nooshin Davoudi
Leila Tahmasbi
Mojgan Bandehpour
Golnaz Asaadi-Tehrani
Department of Biology, Science and Research Branch
West Tehran Islamic Azad University [Tehran] (WTIAU)
Cellular and Molecular Biology Research Center (SBU)
Shahid Beheshti University of Medical Sciences [Tehran] (SBUMS)
Shahid Beheshti University-Shahid Beheshti University
Institut Pasteur d'Iran
Réseau International des Instituts Pasteur (RIIP)
Department of Hematology
Shahid Beheshti University-Shahid Beheshti University-Faculty of Paramedical Sciences
Biotechnology Department
Faculty of Medicine-Shahid Beheshti University of Medical Sciences [Tehran] (SBUMS)
SinaMirzaahmadi
Golnaz Asaadi-Tehrani
Mojgan Bandehpour
Nooshin Davoudi
Leila Tahmasbi
Nahid Hosseinzadeh
Hasan Mirzahoseini
Kazem Parivar
Bahram Kazemi
Faculty of Paramedical Sciences-Shahid Beheshti University of Medical Sciences [Tehran] (SBUMS)
Source :
Journal of Biomedicine and Biotechnology, Journal of Biomedicine and Biotechnology, Hindawi Publishing Corporation, 2011, 2011, pp.873874. ⟨10.1155/2011/873874⟩, Journal of Biomedicine and Biotechnology, Vol 2011 (2011)
Publication Year :
2011
Publisher :
Hindawi Limited, 2011.

Abstract

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 LizardLeishmania, a trypanosomatid protozoan as a host for expression of FVII. Plasmid containing cDNA encoding full-length human FVII was introduced into LizardLeishmaniaand 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, thatLeishmaniacells can be used as an expression system for producing recombinant FVII.

Details

ISSN :
11107251 and 11107243
Volume :
2011
Database :
OpenAIRE
Journal :
Journal of Biomedicine and Biotechnology
Accession number :
edsair.doi.dedup.....55babc4974e7ffcc2f5dfe644cd33612