Construction of the Recombinant Lentiviral Vector Containing Human GH1 Gene and Its Expression in HEK293T Cells

Document Type : Research Articles

Authors

• Zahra Roudbari ¹
• Mohammadreza Nassiri ²
• Mojtaba Tahmoorespur ²
• Aliakbar Haddad-Mashadrizeh ²
• Ali Javadmanesh ²

¹ Ferdowsi University of MashhadUniversity of Mashhad

² Ferdowsi University of Mashhad

https://doi.org/10.22067/jcmr.v8i2.22857

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
• Production protein
• HEK 293 cells

20.1001.1.20089147.2016.8.2.4.4

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