Transfection of mouse PPARgamma1 cDNA in to the bovine fibroblast cells and analysis of its intracellular expression using EGFP gene reporter

Subject Areas : Journal of Large Animal Clinical Science Research(JLACSR)

Received: 2021-07-10 Accepted : 2021-07-10 Published : 2009-03-21

Keywords: cloning, Fibroblast, PPARγ1, transfection,

Abstract :

The main aim of this research is to study nuclear localization of mouse PPARγ1 cDNA into pEGFP-C1. To investigate the nuclear localization of PPARγ1 protein linked to EGFP marker gene into bovine fibroblast and analysis of intracellular green fluorescency Mouse PPARγ1 cDNA in a mammalian expression vector (pEGFP-C1) in a chimeric cDNA type, encompassing PPARγ1 with EGFP cDNA. EGFP marker was used for monitoring. To confirm the intracellular localization of EGFP- PPARγ1, bovine fibroblast cells were transfected with the 2.4 µg constructed plasmid and 6 µl Lipofectamine 2000. The related product was entered into the nucleus of bovine fibroblasts after transfection of its cDNA. As expected, chimeric cDNA of EGFP-PPARγ1 correctly expressed and related protein was dominantly entered to the nuclei of the cells. Thus the recombinant plasmid could be applicable for further studies to unravel the further aspects of PPARγ1 function. Moreover fusion of EGFP with PPARγ1 does not hamper the nuclear targeting of PPARγ1.  

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