Intracellular Localization of FLAG-Peroxisomal Protein in Chinese Hamster Ovary (CHO) Cells

نویسندگان

  • Farzaneh Rabiee Department of Stem Cell, Royan Institute, Isfahan Research Campus, P.O. Box 81589-68433, Isfahan, I.R. Iran
  • Hossein Baharvand Cell Sciences Research Center, Royan Institute, P.O. Box 19395-4644, Tehran, I.R. Iran
  • Kamran Ghaedi Department of Biology, School of Sciences, University of Isfahan, P.O. Box 81746-73441, Isfahan, I.R. Iran
  • Khadijeh Karbalaei Department of Stem Cell, Royan Institute, Isfahan Research Campus, P.O. Box 81589-68433, Isfahan, I.R. Iran
  • Malihe Nazari Jahantigh Department of Biology, School of Sciences, University of Isfahan, P.O. Box 81746-73441, Isfahan, I.R. Iran
  • Maryam Ostad Sharif Department of Basic Medical Sciences, School of Dentistry, Khorasgan Branch, Islamic Azad University, Isfahan, I.R. Iran
  • Marzieh Nematollahi Department of Stem Cell, Royan Institute, Isfahan Research Campus, P.O. Box 81589-68433, Isfahan, I.R. Iran
  • Mehran Miroliaei Department of Biology, School of Sciences, University of Isfahan, P.O. Box 81746-73441, Isfahan, I.R. Iran
  • Mohamad Hossein Nasr Isfahani Department of Stem Cell, Royan Institute, Isfahan Research Campus, P.O. Box 81589-68433, Isfahan, I.R. Iran
  • Shahnaz Razavi Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, P.O. Box 81746-73471, Isfahan, I.R. Iran
  • Somayeh Tanhaei Department of Stem Cell, Royan Institute, Isfahan Research Campus, P.O. Box 81589-68433, Isfahan, I.R. Iran
چکیده مقاله:

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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intracellular localization of flag-peroxisomal protein in chinese hamster ovary (cho) cells

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عنوان ژورنال

دوره 6  شماره 3

صفحات  174- 180

تاریخ انتشار 2008-07-01

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