多浪羊BMP7基因克隆、生物信息学分析及其在不同初情期阶段组织表达分析

doi:10.16431/j.cnki.1671-7236.2023.07.004

Abstract

Abstract: 【Objective】 The sequence of bone morphogenetic protein-7 (BMP7) gene in Dolang sheep was cloned and bioinformatics analyzed, and the expression patterns of five tissues in three stages of puberty were detected, which provided reference for exploring the biological function of BMP7 gene in Dolang sheep during the puberty.【Method】 The sequence of BMP7 gene in Dolang sheep was amplified by PCR, and cloned and sequenced, the phylogenetic tree was constructed using Mega 7.0 software.The physicochemical properties and structural function of BMP7 protein was analyzed by bioinformatics software.The expression of BMP7 gene in hypothalamus, pituitary gland, ovary, oviduct and uterus of Dolang sheep at prepuberty, puberty and postpuberty were detected by Real-time quantitative PCR.【Result】 The BMP7 gene sequence of Dolang sheep was successfully cloned, which was 1 334 bp long, and the CDS sequence was 1 296 bp which encoded 431 amino acids.The BMP7 gene sequence of Dolang sheep had the closer genetic relationship with Ovis aries and Bos taurus, followed by Homo sapiens, Macaca mulatta and Equus caballus, and had the furthest genetic relationship with Mus musculus and Canis lupus familiaris.Bioinformatics analysis results showed that the molecular formula of BMP7 protein was C₂₂₀₀H₃₃₇₄N₆₃₂O₆₂₈S₁₇, the theoretical isoelectric point (pI) was 8.17, the total number of atoms was 6 851, the instability index was 53.18, the lipid solubility coefficient was 76.96, and the total average hydrophilicity was-0.419.BMP7 protein was an unstable hydrophilic basic protein.The secondary structure was mainly consisted of alpha helix and random coil, and the prediction result of tertiary structure was consistent with the secondary structure.Real-time quantitative PCR results showed that BMP7 gene was expressed in prepuberty, puberty and postpuberty of Dolang sheep, and the expression in hypothalamus was significantly higher than that in other tissues (P<0.05).The expression of BMP7 gene was significantly increased in hypothalamus and uterus from prepuberty to puberty (P<0.05), while decreased significantly from puberty to postpuberty (P<0.05).【Conclusion】 The BMP7 gene sequence of Dolang sheep was cloned successfully, and this gene was expressed in five tissues at three different stages of puberty.The results provided a reference for exploring the role of BMP7 gene in puberty of Dolang sheep.

Key words: Dolang sheep; BMP7 gene; cloning; puberty

CLC Number:

WANG Chenguang, SUI Zhiyuan, ZHANG Yongjie, ZHANG Zhishuai, LI Xiaojun, XING Feng. Cloning and Bioinformatics Analysis of BMP7 Gene and Its Tissue Expression at Different Stages of Puberty in Dolang Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2629-2638.

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Cloning and Bioinformatics Analysis of BMP7 Gene and Its Tissue Expression at Different Stages of Puberty in Dolang Sheep

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