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

ABSTRACT

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.