水貂酪氨酸酶(TYR)基因克隆、SNPs筛查及其皮肤组织mRNA差异表达分析

doi:10.11843/j.issn.0366-6964.2021.007

Abstract

Abstract: The aim of this study was to clone and analyze the complete coding region sequence of TYR gene, further reveal the relationship between SNPs and its mRNA differential expression level in skin and coat color phenotype in mink (Neovison vison). A total of 301 blood samples of 7-month-old male minks (Jinzhou black, Mingwei silverblue and Red eye white) were collected, and the polymerase chain reaction (PCR) method was used to clone 5 exons of the mink TYR gene in sections and the complete coding sequence (CDS) was spliced. The PCR products were directly sequenced to screen SNPs in exon 1, 4 and 5 of TYR gene in minks with three kinds of coat color phenotypes. The back skin tissues of 9 7-month-old male minks (3 black, gray and white coats each) were collected, and quantitative real-time PCR (qRT-PCR) technology was used to detect TYR mRNA expression levels in skin of minks with 3 coat colors. The results showed that the mink TYR gene sequence was 2 391 bp in length, containing 5 complete exons, the full-length coding region was 1 596 bp, which encoded 531 amino acids containing signal peptide(18 amino acids) and mature peptide(513 amino acids). The sequence had been submitted GenBank database and its accession number was KJ716783. SNPs analysis showed that there was no mutation site in exon 4 and 5 of TYR gene, and two mutations were found in exon 1, c.441G>A and c.138T>A. However, c.441G>A was synonymous mutation and existed only in Jinzhou black mink population. The qRT-PCR results showed that TYR mRNA was expressed in the skin of all the three kinds of coat color types of minks, and its expression level in the Jinzhou black mink was extremely significantly higher than that in Mingwei silverblue and Red eye white mink (P<0.01). The TYR mRNA expression level in Mingwei silverblue mink was significantly higher than that in Red eye white mink (P<0.05). The results of present study indicated that the c.138T>A locus and mRNA expression levels of TYR gene might be associated with the coat color phenotype of mink.

Key words: mink, TYR gene, SNPs, coat color trait, differential expression, skin

CLC Number:

S813.1

SONG Xingchao, LIU Linling, PAN Hongjun, ZHAO Jiaping, JIA Yun, YANG Fuhe, XU Chao. Cloning,SNPs Screening and mRNA Differential Expression Analysis of TYR Gene in Skin of Mink(Neovison vison)[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 66-76.

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Cloning,SNPs Screening and mRNA Differential Expression Analysis of TYR Gene in Skin of Mink(Neovison vison)

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