Expression of mouse peroxisome proliferator activated receptor ?_2 in NIH3T3 cells induced by recombinant retrovirus vector / 中华内分泌代谢杂志

Objective To express the mouse peroxisome proliferator activated receptor ? 2 (mPPAR? 2) in NIH3T3 cells induced by the recombinant retrovirus vector. Methods mPPAR? 2 gene digested from the recombinant plasmid pcDNA3/mPPAR? 2 containing the target gene segment was subcloned into retrovirus vector pGCEN to generate the recombinant retrovirus pGCEN/mPPAR? 2. The recombinant retrovirus pGCEN/mPPAR? 2 and pGCEN were used to transfect PA317 cells with LipofectAMINE, and anti G418 clones of PA317 cells were selected and viral supernatants were harvested and used to infect NIH3T3 cells. The expressing products were identified with immune flurescence assay (IFA) and Western Blot. Results The recombinant retrovirus pGCEN/mPPAR? 2 was constructed, and 5?10 4 CFU/ml and 6?10 5 CFU/ml of pGCEN/mPPAR? 2 containing and pGCEN containing viral supernatants were obtained respectively. mPPAR? 2 was expressed in NIH3T3 cells mediated by the recombinant retrovirus vector. Conclusion This work is the basis for the foundation of adipocyte differentiation model in vitro and further researching on the molecular mechanism of adipocyte differentiation induced by PPAR? 2.

Molecular cloning of mouse peroxisome proliferator activated receptor ?_2 and characterization of its expressing product in eukaryotic cells / 中华内分泌代谢杂志

Objective To provide an approach to research of peroxisome proliferator activated receptor (PPAR) ? 2 function, mouse PPAR? 2 (mPPAR? 2) gene was cloned and its transient expression in eukaryotic cells was carried out. Methods mPPAR? 2 mRNA from epididymis fat pad of Chinese Kunming mice was amplified by RT PCR and subcloned into plasmid pcDNA3 to generate the recombinant plasmid pcDNA3/mPPAR? 2 which was confirmed to contain the amplified target gene segments with fluorescence sequencing. The recombinant plasmid pcDNA3/mPPAR? 2 was used to transfect COS 7 with lipofectamine and the expressing product was detected with immune fluorescence assay and Western blot. Results The sequencing results for amplified target gene showed that the sequence of mPPAR? 2 from epididymis fat pad of Chinese Kunming mice is similar to that of mouse PPAR? 2 in Genbank, only at the site of 383 amino acid where Ser (AGC) substitutes Asn (AAC). pcDNA3/mPPAR? 2 was efficiently expressed in eukaryotic cells in vitro. Conclusion This work is the experimental basis for further researching on PPAR? 2 function.