intranuclear localization of egfp-mouse pparγ1 in bovine fibroblast cells
نویسندگان
چکیده
background: the aim of this study was to clone pparγ1 cdna in an appropriate mammalian expression vector, with a chimeric cdna form, encompassing pparγ with enhanced green fluorescent protein (egfp) cdna. this recombinant plasmid will be used for further analyses to investigate the molecular mechanism of pparγ1 for neural differentiation process. moreover, the nuclear localization of the pparγ1 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, pparγ1 cdna was synthesized and amplified to produce the entire length of orf. rt-pcr products containing pparγ1 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 pparγ1 cdna were selected for plasmid preparations and additional analysis was performed to ensure that pparγ1 cdna was inserted properly. finally, to confirm the intracellular localization of egfp-pparγ1, bovine fibroblast cells were transfected with the recombinant plasmid. results: our results from enzymatic digestion and sequencing confirmed, as expected, that pparγ1 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: pparγ1 cdna was cloned and sorted into nuclear compartments of bovine fibroblast cells upon transfection.
منابع مشابه
I-11: Dedifferentiation of Mouse Fibroblast Cells by Chemical Induction
Induced pluripotent stem cells (iPSCs) generated by ectopic expression of four transcription factors have great promises for regenerative medicine in humans. Since the initial report of iPSCs by viral transfection, ample efforts have been made in the generation of iPSCs through nonviral approaches. Small molecules offer the advantages of low cost without genomic modification and have been used ...
متن کاملthe effect of mouse embryonic fibroblast in direct differentiation of mouse embryonic stem cells
background: since embryonic stem (es) cells have the dual ability to proliferate indefinitely and differentiate into multiple tissue types, es cells could potentially provide an unlimited cell supply for human transplantation. objective: in order to study the differentiation of mouse embryonic stem (mes) cells, they were cultured in suspension by using es media without leukemia inhibitory facto...
متن کاملIntranuclear localization of snRNP antigens
Anti-Sm antibodies recognize a group of small, nuclear RNA-protein complexes (snRNPs) containing U1, U2, U4, U5, and U6 snRNAs. Anti-RNP antibodies only react with U1 snRNA-containing complexes. The intranuclear distribution of snRNP particles was studied by double immunofluorescence staining of human fibroblasts. Mouse monoclonal anti-Sm antibodies and polyclonal patient sera reacting with dif...
متن کاملEvaluation of Chronotropic Properties of Mouse Embryonic Stem Cells-Derived Cardiomyocytes After Fibroblast Growth Factor Treatment
Purpose: We investigated the effect of (bFGF) (basic-Fibroblast Growth Factor) on the differentiation of divided cardiomyocytes from mouse embryonic stem cells (ES) and their pharmacological properties. Materials and Methods: The mouse embryonic stem cells (Royan B1) were cultured as 800 cells per 20µl of a hanging drop. After two days, ES cells in each drop aggregated to form embryoid bodies ...
متن کاملImprovement of Expression of α6 and β1 Integrins by the Co-culture of Adult Mouse Spermatogonial Stem Cells with SIM Mouse Embryonic Fibroblast Cells (STO) and Growth Factors
متن کامل
The role of fibroblast growth factor receptor 2 (FGFR2) in differentiation of bovine spermatogonial stem cells (SSCs)
The receptors 1 and 2 of fibroblast growth factor (FGFR1 and FGFR2, respectively) have been observed in all types of testicular cells. Culture on extracellular matrix (ECM) has been observed to lead to initiation of differentiation in spermatogonial stem cells (SSCs). The present study was carried out to investigate whether FGFR1 and FGFR2 play a role in SSCs differentiation. Following isolatio...
متن کاملمنابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
cell journalجلد ۱۲، شماره ۱، صفحات ۹۷-۱۰۴
میزبانی شده توسط پلتفرم ابری doprax.com
copyright © 2015-2023