努比亚山羊Myf6基因的克隆及组织表达研究

doi:10.16431/j.cnki.1671-7236.2021.11.018

Abstract

Abstract: The research was aimed to clone myogenic factor 6 (Myf6) gene in Nubian goat and conducted bioinformatics analysis on it. The expression difference of Myf6 gene in different tissues of Nubian goat and the expression difference of longissimus muscle tissue of Nubian goat and Longlin goat were detected. Using the cDNA from the longissimus dorsi muscle tissue of Nubian goat as a template, PCR amplified the complete coding region (CDS) of the Myf6 gene and then verified the bacterial solution. Sequence similarity alignment and phylogenetic tree construction were carried out. The physicochemical properties of Myf6 protein were analyzed and its hydrophilicity/hydrophobicity was predicted. The protein secondary structure and tertiary structure were predicted. The expression of Myf6 gene in heart, liver, spleen, kidney, longissimus dorsi muscle, subcutaneous fat, rumen and Longlin goat longissimus dorsi muscle were detected by Real-time quantitative PCR. The results showed that the CDS sequence of Myf6 gene in Nubian goat was 729 bp and encoded 242 amino acids. The similarity of Myf6 amino acid sequence between Nubian goat and Ovis aries, Bos taurus, Sus scrofa, Equus asinus, Homo sapiens and Mus musculus were 99.3%, 98.8%, 94.1%, 93.8%, 93.4% and 89.0%, respectively. The analysis of the physical and chemical properties of the protein showed that the molecular weight of the goat Myf6 protein was 26.98 ku, which was an unstable acidic protein. The hydrophilic and hydrophobic analysis found that the protein was a hydrophilic protein. The transmembrane structure and signal peptide prediction results showed that the protein did not contain transmembrane structure and signal peptide, and was not a transmembrane protein and secreted protein. The prediction results of secondary structure showed that the protein was mainly composed of random coil (49.59%), followed by alpha helix (33.88%), extended chain (9.50%) and beta turn (7.02%). The prediction results of the tertiary structure were consistent with those of the secondary structure. The results of Real-time quantitative PCR showed that the expression of Myf6 gene in the longissimus dorsi muscle of Nubian goat was the highest, significantly higher than that of other tissues (P<0.05), and the expression in the kidney was the lowest, but there was no significant difference with heart, liver, spleen, subcutaneous fat and rumen (P>0.05). Compared the expression of Myf6 gene in the longissimus dorsi muscle of Nubian goat and Longlin goat, the expression level of Nubian goat was significantly higher than that of Longlin goat (P<0.05). The experiment provided a reference for further revealing the role of Myf6 gene in muscle differentiation.

Key words: goat; Myf6 gene; cloning; tissue expression

CLC Number:

S827

SONG Ying, ZHANG Sanbao, SHEN Yujian, HU Yan, ZHANG Yu, ZOU Juhong, XU Jianjian, LIAN Zitong, ZOU Jianwei, WEI Yingming, ZHENG Zihua, JIANG Qinyang. Cloning and Tissue Expression of Myf6 Gene in Nubian Goat[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4084-4093.

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Cloning and Tissue Expression of Myf6 Gene in Nubian Goat

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