As a resource for FVII gene expression, a variety of recombinant protein expression systems have been developed. The authors used a novel protein expression system based on the Iranian Lizard Leishmania, a trypanosomatid protozoan, as a host for FVII expression in the current study. The synthesis of recombinant activated FVII (rFVIIa) was considered based on an understanding of the role of activated FVII in the treatment of coagulation disorders. Lizard Leishmania was infected with a plasmid containing cDNA encoding full-length human FVII, and positive transfectants were analyzed using SDS. PAGE and Western blot tests were performed. Furthermore, the PT assay was used to determine the biological activity of purified protein. The expression of FVII at the mRNA and protein levels was examined in a recombinant strain containing a construct. Purified rFVII was obtained, and it was confirmed that the purified compound was indeed rFVII. Western blot analysis was performed. With the purified rFVII, the clotting time in the PT assay was reduced by about 30 seconds. Finally, for the first time, this study demonstrated that Leishmania cells can be used as an expression system for producing recombinant FVII.
Author (S) Details
Dr. Sina Mirzaahmadi
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Golnaz Asaadi-Tehrani
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Dr . Mojgan Bandehpour
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Nooshin Davoudi
Biotechnology Department, Pasteur Institute, Tehran, Iran.
Leila Tahmasbi
Department of Hematology, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Nahid Hosseinzadeh
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Hasan Mirzahoseini
Biotechnology Department, Pasteur Institute, Tehran, Iran.
Kazem Parivar
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Dr . Bahram Kazemi
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran and Biotechnology Department, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran and Parasitology Department, Schooll of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
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