INTRANUCLEAR LOCALIZATION OF EGFP-MOUSE PPARγ1 IN BOVINE FIBROBLAST CELLS

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GHASEMI SORAYA; GHAEDI K.; NASRESFAHANI MOHAMMAD HOSSEIN; TANHAEI S.; RABIEI F.; KARBALAEI KH.; BAHARVAND HOSSEIN; ESMAEILI A.A.GH.

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Journal: Cell Journal (Yakhteh) Year:2010 | Volume:12 | Issue:1 (45) Page(s): 97-104

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Title

INTRANUCLEAR LOCALIZATION OF EGFP-MOUSE PPARγ1 IN BOVINE FIBROBLAST CELLS

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Keywords

PPARγQ1

NUCLEAR TARGETINGQ1

ENHANCED GREEN FLUORESCENT PROTEINQ1

CLONINGQ1

TRANSFECTIONQ1

Abstract

Objective: The aim of this study was to clone PPARg1 cDNA in an appropriate mammalian expression vector, with a chimeric cDNA form, encompassing PPARg with ENHANCED GREEN FLUORESCENT PROTEIN (EGFP) cDNA. This recombinant plasmid will be used for further analyses to investigate the molecular mechanism of PPARg1 for neural differentiation process. Moreover, the nuclear localization of the PPARg1 protein linked to EGFP marker was chased by using transient TRANSFECTION of a constructed plasmid into bovine fibroblast cells.Materials and Methods: Total RNA was extracted from the fatty tissue of an adult mouse. Using specific pair primers, PPARg1 cDNA was synthesized and amplified to produce the entire length of ORF. RT-PCR products containing PPARg1 cDNA were treated by enzymatic digestion and inserted into the pEGFP-C1 downstream from EGFP cDNA. The constructed vector was used for transformation into bacterial competent cells. Positive colonies which showed inserted PPARg1 cDNA were selected for plasmid preparations and additional analysis was performed to ensure that PPARg1 cDNA was inserted properly. Finally, to confirm the intracellular localization of EGFP-PPARg1, bovine fibroblast cells were transfected with the recombinant plasmid.Results: Our results from enzymatic digestion and sequencing confirmed, as expected, that PPARg1 cDNA was amplified and cloned correctly. This cDNA gene encompassed 1428 bp. The related product was entered into the nucleus of bovine fibroblasts after TRANSFECTION of its cDNA.Conclusion: PPARg1 cDNA was cloned and sorted into nuclear compartments of bovine fibroblast cells upon TRANSFECTION.

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APA: Copy

GHASEMI, SORAYA, GHAEDI, K., NASRESFAHANI, MOHAMMAD HOSSEIN, TANHAEI, S., RABIEI, F., KARBALAEI, KH., BAHARVAND, HOSSEIN, & ESMAEILI, A.A.GH.. (2010). INTRANUCLEAR LOCALIZATION OF EGFP-MOUSE PPARγ1 IN BOVINE FIBROBLAST CELLS. CELL JOURNAL (YAKHTEH), 12(1 (45)), 97-104. SID. https://sid.ir/paper/16386/en

Vancouver: Copy

GHASEMI SORAYA, GHAEDI K., NASRESFAHANI MOHAMMAD HOSSEIN, TANHAEI S., RABIEI F., KARBALAEI KH., BAHARVAND HOSSEIN, ESMAEILI A.A.GH.. INTRANUCLEAR LOCALIZATION OF EGFP-MOUSE PPARγ1 IN BOVINE FIBROBLAST CELLS. CELL JOURNAL (YAKHTEH)[Internet]. 2010;12(1 (45)):97-104. Available from: https://sid.ir/paper/16386/en

IEEE: Copy

SORAYA GHASEMI, K. GHAEDI, MOHAMMAD HOSSEIN NASRESFAHANI, S. TANHAEI, F. RABIEI, KH. KARBALAEI, HOSSEIN BAHARVAND, and A.A.GH. ESMAEILI, “INTRANUCLEAR LOCALIZATION OF EGFP-MOUSE PPARγ1 IN BOVINE FIBROBLAST CELLS,” CELL JOURNAL (YAKHTEH), vol. 12, no. 1 (45), pp. 97–104, 2010, [Online]. Available: https://sid.ir/paper/16386/en

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