Cloning of gene PPARγfrom Guangxi Bama mini-pig and tissue expression difference analysis for it

ObjectiveIn this study,peroxisome proliferator-activated receptorγgene(PPARγ)from Guangxi Bama mini-pig was cloned and its expressions in different tissues were determined in order to lay a foundation for further study-ing the mechanism of gene PPARγ in mycoplasmal pneumoniae of swine(MPS).MethodGene PPARγ from Guangxi Bama mini-pig was cloned by RT-PCR,and BLAST comparison analysis and bioinformatics analysis on its deduced pro-tein were conducted. The expressions of gene PPARγ were detected through real-time fluorescence quantitative PCR in heart,spleen,lung,kidney,large intestine,small intestine,muscle and fat of Guangxi Bama mini-pig of 18 months.ResultThe results showed that open reading frame of PPARγgene from Guangxi Bama mini-pig was 1515 bp in length, encoding 504 amino acids. The homology between the deduced amino acid sequence of gene PPARγfrom Guangxi Bama mini-pig and the published porcine gene PPARγ(FJ436399.1)in GenBank were the highest,reaching 100.0%;Guangxi Bama mini-pig shared 92.9%,91.8%,91.5%,90.5%and 83.5%homology with amino acid sequences from Bos taurus(NM_181024.2),Homo sapiens(NM_005037.5),Macaca mulatta(NM_001032860.1),Mus musculus(NM_001308354.1), and Anser cygnoides(KJ019822.1) respectively,which indicated that PPARγ was highly conservative. The molecular weight of protein PPARγ was 57380.91 Da,its theoretical isoelectric point(pI)was 6.88,coefficient of instability was 49.26,fat index was 88.21,hydrophilic average was-0.293,which indicated that PPARγwas hydrophilic protein. The secondary structure was mainly alpha helix,accounting for 39.09%of the total. The expression of gene PPARγin large in-testine of Guangxi Bama mini-pig was the highest,extremely significantly higher than those in other tissues(P<0.01), while its expression was very low in kidney,heart and muscle.ConclusionGene PPARγ expresses in all tissues of Guangxi Bama mini-pig of 18 months,but the expression shows significant difference in different tissues,which may be related to its functional differences in various tissues.