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

Authors

  • 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
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.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

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

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 construc...

full text

EXPRESSION OF HUMAN PROTEINASE 3 IN CHINESE HAMSTER OVARY CELLS (CHO-CELLS)

Proteinase 3(PR3) is a human polymorphonuclear leukocyte serine proteinase and is the main target antigen for antineutrophil cytoplasmic antibodies (ANCA) found in Wegener's granulomatosis (WG). We developed a stable expression system for conformationally intact recombinant PR3 (rPR3) in Chinese hamster ovary cells (CHO-cells). The part of PR3 cDNA that encoded the active form of PR3 was s...

full text

Cloning and Expression of Recombinant Human Interleukin-7 in Chinese Hamster Ovary (CHO) Cells

Background: The critical role of interleukin-7 (IL-7) in homeostatic proliferation and T cell survival has made it a promising cytokine for the treatment of various clinical conditions, especially those associated with lymphopenia. Methods: In the present study we expressed recombinant human interleukin-7 (rhIL-7) in Chinese hamster ovary (CHO)-K1 cells. CHO-K1 cells were stably transfected ...

full text

Using transient transfected Chinese hamster ovary (CHO) cells by pET28a-LIC-NTF4 in gene therapy

Introduction: Neurotrophins, as a family of proteins, are responsible for induction of the survival, development, and function of neurons. Also, neurotropic factors are growth factors like Neurotrophins that help neurons survive. Moreover, Neurotrophins differentiate between progenitor cells so that neurons are formed. Despite the fact that the majority of mammalian brain neurons are produced p...

full text

expression of human proteinase 3 in chinese hamster ovary cells (cho-cells)

proteinase 3(pr3) is a human polymorphonuclear leukocyte serine proteinase and is the main target antigen for antineutrophil cytoplasmic antibodies (anca) found in wegener's granulomatosis (wg). we developed a stable expression system for conformationally intact recombinant pr3 (rpr3) in chinese hamster ovary cells (cho-cells). the part of pr3 cdna that encoded the active form of pr3 was s...

full text

Molecular Cloning and Expression of Human Gamma Interferon (IFN-g) Full cDNA in Chinese Hamster Ovary (CHO) Cells

Background: IFN-g is mostly secreted by activated CD4+ , CD8+ T cells and NK cells. This cytokine has immunomodulatory, anti-cancer and anti-microbial effects and is important for prophylaxis, diagnosis and treatment of chronic infections and cancers. Objective: The purpose of this study was to clone the full cDNA of human IFN-g and express it in CHO cell line. Methods: Lymphocytes from a healt...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 6  issue 3

pages  174- 180

publication date 2008-07-01

{@ msg @}

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023