Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression System

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 conta...

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Main Authors: Sina Mirzaahmadi, Golnaz Asaadi-Tehrani, Mojgan Bandehpour, Nooshin Davoudi, Leila Tahmasbi, Nahid Hosseinzadeh, Hasan Mirzahoseini, Kazem Parivar, Bahram Kazemi
Format: Article
Language:English
Published: Hindawi Limited 2011-01-01
Series:Journal of Biomedicine and Biotechnology
Online Access:http://dx.doi.org/10.1155/2011/873874
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spelling doaj-1e492d296f6f45858225a5326cdbb9fb2020-11-25T01:38:03ZengHindawi LimitedJournal of Biomedicine and Biotechnology1110-72431110-72512011-01-01201110.1155/2011/873874873874Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression SystemSina Mirzaahmadi0Golnaz Asaadi-Tehrani1Mojgan Bandehpour2Nooshin Davoudi3Leila Tahmasbi4Nahid Hosseinzadeh5Hasan Mirzahoseini6Kazem Parivar7Bahram Kazemi8Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranDepartment of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IranBiotechnology Department, Pasteur Institute, Tehran, IranDepartment of Hematology, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, IranCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IranBiotechnology Department, Pasteur Institute, Tehran, IranDepartment of Biology, Science and Research Branch, Islamic Azad University, Tehran, IranCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IranThe 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.http://dx.doi.org/10.1155/2011/873874
collection DOAJ
language English
format Article
sources DOAJ
author Sina Mirzaahmadi
Golnaz Asaadi-Tehrani
Mojgan Bandehpour
Nooshin Davoudi
Leila Tahmasbi
Nahid Hosseinzadeh
Hasan Mirzahoseini
Kazem Parivar
Bahram Kazemi
spellingShingle Sina Mirzaahmadi
Golnaz Asaadi-Tehrani
Mojgan Bandehpour
Nooshin Davoudi
Leila Tahmasbi
Nahid Hosseinzadeh
Hasan Mirzahoseini
Kazem Parivar
Bahram Kazemi
Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression System
Journal of Biomedicine and Biotechnology
author_facet Sina Mirzaahmadi
Golnaz Asaadi-Tehrani
Mojgan Bandehpour
Nooshin Davoudi
Leila Tahmasbi
Nahid Hosseinzadeh
Hasan Mirzahoseini
Kazem Parivar
Bahram Kazemi
author_sort Sina Mirzaahmadi
title Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression System
title_short Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression System
title_full Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression System
title_fullStr Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression System
title_full_unstemmed Expression of Recombinant Human Coagulation Factor VII by the Lizard Leishmania Expression System
title_sort expression of recombinant human coagulation factor vii by the lizard leishmania expression system
publisher Hindawi Limited
series Journal of Biomedicine and Biotechnology
issn 1110-7243
1110-7251
publishDate 2011-01-01
description 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.
url http://dx.doi.org/10.1155/2011/873874
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