Author/Authors :
Roudbari، Zahra نويسنده Faculty of Agriculture,Department of Animal Science,Ferdowsi University of Mashhad,Mashhad,Iran , , Nassiri، Mohammadreza نويسنده Faculty of Agriculture,Department of Animal Science, Iran 2-Cell and Molecular Biotechnology Research Group,Ferdowsi University of Mashhad, Institute of Biotechnology,Mashhad,Iran , , Tahmoorespur، Mojtaba نويسنده Faculty of Agriculture,Department of Animal Science,Ferdowsi University of Mashhad,Mashhad,Iran , , Haddad-Mashadrizeh، Aliakbar نويسنده Faculty of Science,Cell and Molecular Biotechnology Research Group, Department of Biology,Institute of Biotechnology, Ferdowsi University of Mashhad,Mashhad,Iran , , Javadmanesh، Ali نويسنده Faculty of Agriculture,Department of Animal Science,Ferdowsi University of Mashhad,Mashhad,Iran ,
Abstract :
Human growth hormone (hGH) is a protein with multiple roles in a range of biological functions such as protein, carbohydrates and lipid metabolisms as well as immunity, tissue development and overall growth. One of the major class of biopharmaceuticals in mammalian cells is the production of recombinant pharmaceutical proteins. In this study, we constructed a lentiviral vector carrying coding region of human GH1 (hGH) gene in order to production of recombinant hGH in mammalian cell line. hGH gene was amplified from a plasmid containing full-length hGH coding sequence and then cloned into the lentiviral vector pCDH-GFP. The HEK293T cells were transduced by the lentivirus particles as a targeted cell. hGH expression status in the recombinant cells were confirmed by RT-PCR. Additionally, western blotting analysis results showed that the recombinant cells maintained a stable hGH expression during five weeks of continuous culture. In conclusion, results of current study suggested that constructed lentiviral vector can potentially be used for a stable production of recombinant hGH protein in HEK293T cells. This methodology could be served as a foundation for further research and may open new insights toward therapeutic protein manufacturing.
Keywords :
hGH , Recombinant lentivirus , HEK 293 cells , Production protein