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Intracellular Localization of Flag-Peroxisomal Protein in Chinese Hamster Ovary (Cho) Cells



Jahantigh MN1 ; Ghaedi K1, 2 ; Isfahani MHN2 ; Tanhaei S2 ; Rabiee F2 ; Karbalaei K2 ; Sharif MO2, 3 ; Nematollahi M2 ; Baharvand H4 ; Razavi S5 ; Miroliaei M1
Authors
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Authors Affiliations
  1. 1. Department of Biology, School of Sciences, University of Isfahan, Isfahan, P.O. Box 81746-73441, Iran
  2. 2. Department of Stem Cell, Royan Institute, Isfahan Research Campus, Isfahan, P.O. Box 81589-68433, Iran
  3. 3. Department of Basic Medical Sciences, School of Dentistry, Khorasgan Branch, Islamic Azad University, Isfahan, Iran
  4. 4. Cell Sciences Research Center, Royan Institute, Tehran, P.O. Box 19395-4644, Iran
  5. 5. Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, P.O. Box 81746-73471, Iran

Source: Iranian Journal of Biotechnology Published:2008

Abstract

Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The aim of this study was to sub-clone the peroxisomal protein (PEP) cDNA into a mammalian expression vector leading to the formation of a chimeric PEP-cDNA containing the FLAG epitope. The FLAG-PEP recombinant cDNA was constructed using the method of splicing by overlap extension polymerase chain reaction (SOE PCR) and inserted into the pUcD2SRaMCSHyg eukaryotic expression vector. To investigate the intracellular localization of the PEP protein that was linked to the FLAG tandem, the constructed plasmid was used for transient transfection of the Chinese hamster Ovary (CHO) cells. The CHO cells that were transfected with the recombinant plasmid showed peroxisomal localization of FLAG-PEP as was previously shown for catalase.
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