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

  • سال انتشار: 1395
  • محل انتشار: مجله سلول و تحقیقات مولکولی، دوره: 8، شماره: 2
  • کد COI اختصاصی: JR_JCMR-8-2_005
  • زبان مقاله: انگلیسی
  • تعداد مشاهده: 599
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نویسندگان

Zahra Roudbari

Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

Mohammadreza Nassiri

Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

Mojtaba Tahmoorespur

Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

Aliakbar Haddad-Mashadrizeh

Cell and Molecular Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad,Iran

چکیده

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 thisstudy, 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

کلیدواژه ها

hGH, Recombinant lentivirus, Production protein, HEK 293 cells

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