延边牛PLIN2基因CDS区克隆、生物信息学分析及其表达规律研究

doi:10.16431/j.cnki.1671-7236.2021.03.006

Abstract

Abstract: The present study was aimed to clone the complete CDS region of Yanbian cattle adipose differentiation-related protein PLIN2 gene,analyze the sequence and basic protein characteristics of PLIN2 gene CDS region through bioinformatics,and explore its presence in different tissues of Yanbian cattle.The 18-month-old castrated Yanbian cattle were selected as the research object.PLIN2 gene of Yanbian cattle was obtained by RT-PCR and gene cloning,and the homology comparison with other species was carried out and the phylogenetic tree was constructed.The PLIN2 encoded protein was predicted by bioinformatics methods the physicochemical properties,potential phosphorylation sites,glycosylation sites,disulfide bond analysis,signal peptides,subcellular localization and high-level structure of the protein,Real-time quantitative PCR was used to detect the expression level of PLIN2 gene in different tissues of Yanbian cattle.The results showed that the CDS of PLIN2 gene in Yanbian cattle was 1 353 bp long and encoded 450 amino acids.The homology comparison showed that Yanbian cattle PLIN2 gene had 100% homology with cattle,98.7% for buffalo,96.7% for sheep,97.1% for goat,85.5% for wild boar,86.3% for human,47.3% for mouse,and 49.1% for pheasant.The phylogenetic tree showed that Yanbian cattle had the closest genetic relationship with Yellow cattle and buffalo,and the farthest genetic relationship with pheasants.PLIN2 protein was an unstable protein with a certain degree of hydrophilicity.There were 61 potential phosphorylation sites,16 potential O-glycosylation sites,12 potential N-glycosylation sites,and 2 disulfide sites,there was no signal peptide,mainly distributed in the nucleus,a small amount in the cytoplasm and mitochondria.The high-level structure of PLIN2 protein predicted that the protein mainly contains a mixed protein composed of α-helix,extended chain,random coils and β-turns,which were connected by random coils,mainly α-helix.Real-time quantitative PCR results showed that PLIN2 gene was expressed the highest in small intestine tissue of Yanbian cattle,followed by longissimus dorsi muscle and kidney tissue.The results of this study could provide a reference for further research on the function of PLIN2 gene.

Key words: Yanbian cattle; PLIN2 gene; cloning; bioinformatics analysis; expression

CLC Number:

S823

SUN Jianfu, CUI Yan, ZHANG Junfang, WANG Ying, SUN Bin, LI Qiang, Seong H. Choi, Jong S. Shin, LI Xiangzi. Cloning,Bioinformatics Analysis and Expression of PLIN2 Gene CDS Region in Yanbian Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(3): 829-838.

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Cloning,Bioinformatics Analysis and Expression of PLIN2 Gene CDS Region in Yanbian Cattle

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