藏羊MSTN基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2022.11.021

Abstract

Abstract: 【Objective】 The objective of this study was to clone and bioinformatic analysis of myostatin (MSTN) gene in Tibetan sheep, and investigate its expression in different tissues of Tibetan sheep, so as to provide a referencce for exploring the biological function of MSTN gene in Tibetan sheep.【Method】 Using cDNA of longissimus dorsi muscle of Tibetan sheep as template, the complete CDS region of MSTN gene of Tibetan sheep was cloned and sequenced, and the sequencing results were spliced by SeqMan program, and the assembled sequence was analyzed and identified by BLAST online procedure.Similarity comparison, phylogenetic tree construction and bioformztics analysis were carried out by bioinformatics softwares, and the expression of MSTN gene in different tissues of Tibetan sheep was detected by Real-time quantitative PCR.【Result】 The CDS length of MSTN gene in Tibetan sheep was 1 128 bp and encode 375 amino acids.The amino acid sequence similarity of MSTN gene in Tibetan sheep were 100.0%, 93.4%, 93.4%, 95.2%, 94.4%, 94.2%, 94.4%, 93.1%, 87.8% and 87.5% with Ovis aries, Bos mutus, Bos taurus, Sus scrofa, Macaca mulatta, Homo sapiens, Pan troglodytes, Canis lupus familiaris, Gallus gallus and Maylandia zebra.The results of phylogenetic tree analysis showed that Tibetan sheep was the closest to Ovis aries and the furthest to Maylandia zebra and Gallus gallus. MSTN protein of Tibetan sheep was a hydrophilic secretory protein, and had instability, without transmembrane structure, contains 1 signal peptide, has 31 potential phosphorylation sites and 2 N-glycosylation modification sites, it was mainly distributed in mitochondria and cytoplasm.The secondary structure of MSTN protein was consistent with the prediction of tertiary structure, mainly random coil, followed by α-helix, extended chain and β-turn, which was a mixed protein.Real-time quantitative PCR results showed that MSTN gene was expressed in hypothalamus, pituitary, heart, liver, spleen, lungs, kidneys, rumen, duodenum, triceps brachio, semitendinosus, quadriceps femoris and longissimus dorsi, and the MSTN gene was expressed in different tissues of Tibetan sheep, and the expression of MSTN gene in triceps brachii and semitendinosus muscle was significantly higher than that in lung, hypothalamus, heart, liver, duodenum, rumen and kidney(P<0.05).【Conclusion】 MSTN gene was successfully cloned from Tibetan sheep, and its expression in muscle was higher than that in visceral tissue, which laid a foundation for further study of the regulatory mechanism of MSTN gene on muscle growth and development in Tibetan sheep.

Key words: MSTN gene; cloning; bioinformatics analysis; gene expression

CLC Number:

S826.8

ZHANG Junxia, HE Na, LI Mingming, LYU Cailing, CUI Yajie, LANG Xia, WANG Cailian. Cloning,Bioinformatics and Tissue Expression Analysis of MSTN Gene in Tibetan Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4307-4317.

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Cloning,Bioinformatics and Tissue Expression Analysis of MSTN Gene in Tibetan Sheep

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