Author/Authors :
-، - نويسنده Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, I.R. Iran Ghaedi, Mahboobe , -، - نويسنده Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, I.R. Iran Sahebghadam Lotfi, Abbas , -، - نويسنده Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, P.O. Box 14115-111, Tehran, I.R. Iran Soleimani, Masoud , -، - نويسنده Department of Bacteriology and Virology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran Shamsara, Mehdi , -، - نويسنده School of Pharmacy, Zanjan University of Medical Sciences, P.O. Box 45195-1338, Zanjan, I.R. Iran Arjmand, Sare , -، - نويسنده Department of Bacteriology and Virology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P.O. Box 14965/161, Tehran, I.R. Iran Adibi, Behzad
Abstract :
In this study, in order to facilitate and accelerate the production of eukaryotic protein alpha 1-antitrypsin (AAT) with correct post-translational modifications, a protein production system based on the transduction of CHO and COS-7 cells using lentiviral vectors was developed. Human AAT cDNA was cloned into a replication-defective lentiviral vector. The transgene AAT-Jred chimer was transferred to CHO and COS-7 cell lines using this vector and its expressions were visualized by fluorescent microscopy. The mRNA expression levels of the AAT genes were determined using Revearse Transcriptase-Polymerase Chain Reaction (RT-PCR) and its secretion into the medium by both cell types was determined using ELISA. The results show that by employing a lentiviral vector, efficient genetic loading of CHO and COS-7 cells with the AAT gene was achieved. In conclusion, by using a Lentivirus-based gene delivery system, large amounts of recombinant human AAT protein were expressed in both CHO and COS-7 cell lines. This expression system possesses key properties that ensure its application in the delivery of therapeutic genes into mammalian cultured cells.
