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Intracellular localization of FLAG-peroxisomal protein in Chinese Hamster Ovary (CHO) cells

Nazari Jahantigh Malihe نويسنده , Ghaedi Kamran نويسنده , Nasr Isfahani Mohammad Hossein نويسنده , Tanhaei Somayeh نويسنده , Rabiee Farzaneh نويسنده , Karbalael Khadijeh نويسنده , Ostad Sharif Maryam نويسنده , Nematollahi Marzieh نويسنده , Baharvand Hossein نويسنده , Razavi Shahnaz نويسنده , Miroliaei Mehran نويسنده

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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) eDNA into a mammalian expression vector leading to the formation of a chimeric PEP-eDNA containing the FLAG epitope. The FLAG-PEP recombinant eDNA 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.

1202020