皱皮香猪HAS2基因SNPs的鉴定及生物信息学分析

doi:10.11843/j.issn.0366-6964.2021.012.007

Abstract

Abstract: To explore whether the systemic skin wrinkle of Xiang pig with wrinkled skin (XPZ) was related to the variation of HAS2(hyaluronan synthase 2) gene, all exon regions of HAS2 gene were cloned by specific PCR method with ordinary Xiang pig(XP) as control. The base variation of gene coding region was analyzed by bioinformatics method, and the distribution frequencies of variant SNP sites in population were detected. Four SNP sites were detected from HAS2 gene of Xiang pig with wrinkled skin,including T221A missense mutation in exon 1 (resulting in leucine substitution to lysine) and A228G synonymous mutation,T183C synonymous mutation in exon 2 and C537T synonymous mutation in exon 4. Bioinformatics analysis showed that 4 haplotypes were formed at T221A and A228G loci. Among them, T-A was the main haplotype (27.5%) and closely linked(r²=0.253) in Xiang pig population with wrinkled skin. Moreover, mutations in T221A and A228G could change HAS2 mRNA free energy and protein structure, and the free energy of TA, TG, AG and AA combinations were -573.29, -580.89, -572.66, -571.03 kJ·mol^-1,respectively. After leucine was replaced by lysine resulted from T221A missense mutation, the percentage of α-helix decreased from 35.52% to 32.97%, and the free crimp increased from 43.17% to 44.51%. Two SNPsites of loci T221A and A228G showed abundant polymorphism in Xiang pig population with 3 genotypes. And both frequencies of A allele in T221A(P<0.05) and G allele in A228G(P<0.01) were significantly higher in Xiang pig with wrinkled skin than that in common Xiang pig. There were no significant difference in the frequencies of C allele in T183C and T allele in C537T between Xiang pig with wrinkled skin and common Xiang pig. The results showed that the two base mutations in exon 1 of HAS2 gene might affect the stability of post-transcriptional mRNA, the three-dimensional structure of protein and HA synthesis, and have a relationship to the thickening and wrinkling of skin in Xiang pig with wrinkled skin.

Key words: Xiang pig with wrinkled skin, hyaluronidase, SNPs, genotype, skin wrinkles

CLC Number:

S828.2

WU Xingdiao, SHI Daohong, HUANG Shihui, NIU Xi, LI Sheng, WANG Jiafu, RAN Xueqin. SNPs Identification and Bioinformatics Analysis of the Hyaluronan Synthase 2 Gene in Xiang Pig with Wrinkled Skin[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3390-3402.

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SNPs Identification and Bioinformatics Analysis of the Hyaluronan Synthase 2 Gene in Xiang Pig with Wrinkled Skin

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