China Animal Husbandry and Veterinary Medicine ›› 2022, Vol. 49 ›› Issue (12): 4513-4523.doi: 10.16431/j.cnki.1671-7236.2022.12.001

• Biotechnology • Previous Articles     Next Articles

Cloning, Bioinformatics and Expression Analysis of DJ-1 Gene in Large White×Landrace Pigs

CHUI Linya1,2, XIA Boce2, HUANG Jiaojiao1   

  1. 1. College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China;
    2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:2022-05-07 Online:2022-12-05 Published:2022-12-01

Abstract: 【Objective】 The purpose of this study was to clone the CDS region of DJ-1 gene in Large White×Landrace pigs and analyzed by bioinformatics, explore the expression pattern of DJ-1 gene in porcine adipose tissue, and lay a foundation for further studies on the role of DJ-1 gene in porcine adipose deposition.【Method】 The CDS region of DJ-1 gene was amplified by PCR using the cDNA of inguinal subcutaneous adipose tissue in Large White×Landrace pigs as a template, and the bioinformatics analysis of DJ-1 protein was performed using online tools.Real-time quantitative PCR was used to measure the expression of DJ-1 gene in adipose tissue of Large White×Landrace pigs, Large White pigs (lean-type) and Laiwu pigs (obese-type), and the expression differences were compared.【Result】 The length of DJ-1 gene CDS in Large White×Landrace pigs was 570 bp, which coded 189 amino acids.According to the phylogenetic tree, Large White×Landrace pigs was most closely related to Bos taurus and most distantly related to Danio rerio.The molecular weight of DJ-1 protein was 19.94 ku, it was stable, acidic and hydrophilic protein, which had one conserved domain called GAT_1 superfamily Thij.The DJ-1 protein had 21 phosphorylation sites, 4 N-glycosylation modification sites and 1 O-glycosylation modification site, but no signal peptide and transmembrane domain.The secondary structure of DJ-1 protein was composed of alpha helix, extended chain, beta turn and random coil, the proportions were 42.86%, 17.99%, 7.94% and 31.22%, respectively, and the tertiary structure prediction results were consistent with them.Protein interaction analysis revealed that DJ-1 protein interacted with SOD2, NDUFV2, SNCA, BCL2L1 and MAP3K.Real-time quantitative PCR results showed that DJ-1 gene was generally expressed in backfat, inguinal fat and perirenal fat of Large White×Landrace pigs, there were no significant difference among them (P>0.05).The expression of DJ-1 gene in Large White pigs was significantly or extremely significantly lower than that of Laiwu pigs (P<0.05 or P<0.01).【Conclusion】 This study successfully cloned the CDS region of DJ-1 gene in Large White×Landrace pigs, and DJ-1 was a stable, acidic and hydrophilic protein.The expression of DJ-1 gene in Laiwu pigs was significantly or extremely significantly higher than that of Large White pigs. The results provided a theoretical reference for further study the function of porcine DJ-1 gene.

Key words: pigs; DJ-1 gene; bioinformatics analysis; expression; adipose

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