Molecular cloning and characterisation of peroxisome proliferator activated receptor gamma1 (PPARgamma1) cDNA gene from guinea pig (Cavia porcellus): determination of tissue distribution of PPARg in guinea pig

摘要

The coding region of guinea pig peroxisome proliferator activated receptor gamma 1 (gpPPAR gamma 1) cDNA was successfully cloned from adipose tissue by reverse transcription polymerase chain reaction (RT-PCR) using the designated primers based on the conserved regions of the other mammalian PPARgamma 1 sequence. From RT-PCR, a combination of three cDNA fragments that comprised of the full length coding region PPARgamma1 cDNA gene were amplified, with the size of 498, 550 and 557 bp, respectively. All three fragments were then successfully assembled by utilising the internal restriction sites present at the overlapping regions to give rise to the full-length coding region of gpPPARgamma 1 with the size of 1428 bp and consisting of 475 amino acids. Guinea pig PPARgamma 1 is highly conserved with those of other species at protein and nucleotide levels. Gene expression studies showed that gpPPArgamma mRNA was predominantly expressed in adipose tissue followed by lung and spleen. However, at the protein level, PPARgamma was found to be expressed in skeletal muscle.