延边牛PPARγ基因的克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2021.08.002

Abstract

Abstract: The purpose of this study was to clone and analyze the peroxisome proliferator-activated receptor γ(PPARγ) gene of Yanbian cattle, and explore its expression patterns in different tissues of Yanbian cattle.Primers were designed based on the sequence of bovine PPARγ gene in the GenBank database (accession No.:NM_181024.2).Taking 18-month-old Yanbian cattle as the experimental object, the PPARγ gene of Yanbian cattle was cloned by RT-PCR, and BLAST in NCBI was used to carry out homology analysis and construct a phylogenetic tree with other species.Bioinformatics softwares were used to analyze the nucleotide sequence and the physicochemical properties, hydrophobicity, signal peptides, transmembrane domains, subcellular localization, phosphorylation sites, glycosylation sites, secondary structure and tertiary structure of PPARγ protein.Real-time quantitative PCR technology was used to detect the expression level of PPARγ gene in heart, liver, spleen, lung, kidney, small intestine, subcutaneous fat, muscle of Yanbian cattle.The results showed that the CDS region sequence of Yanbian cattle PPARγ gene was 1 620 bp, the sequence and phylogenetic tree analysis showed that it had 99.9% homology with that of yellow cattle and 82.1% homology with chicken.PPARγ gene encoded a protein of 505 amino acids, and the protein had no signal peptide and transmembrane domain, which was hydrophilic protein and mainly distributed in the nucleus and cytoplasm.The main secondary structure of Yanbian cattle were alpha helix and random coil, and there were 53 potential phosphorylation sites and 24 O-glycosylation sites.Real-time quantitative PCR results showed that the expression of Yanbian cattle PPARγ gene in spleen, subcutaneous fat and liver was significantly higher than that in heart (P<0.05), the expression of it in lung, kidney, muscle and small intestine was significantly lower than that in heart (P<0.05).The above results could provide reference for further study of the function of PPARγ gene.

Key words: PPARγ; Yanbian cattle; bioinformatics analysis; gene cloning; Real-time quantitative PCR

CLC Number:

S823

ZHANG Junfang, SUN Jianfu, SUN Bin, TANG Lin, WANG Ying, WANG Enze, CUI Yan, LI Qiang, YAN Changguo, LI Xiangzi. Cloning and Bioinformatics Analysis of PPARγ Gene of Yanbian Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2695-2704.

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Cloning and Bioinformatics Analysis of PPARγ Gene of Yanbian Cattle

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