绵羊MUSTN1基因的克隆、序列分析及其组织表达

doi:10.3969/j.issn.1004-1524.2017.10.10

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (10): 1661-1668.DOI: 10.3969/j.issn.1004-1524.2017.10.10

绵羊MUSTN1基因的克隆、序列分析及其组织表达

高建锋^{1, 2}, 卢曾奎^{1, 2}, 马友记^{1, 2, *}, 李讨讨^{1, 2}, 赵兴绪³

1.甘肃农业大学动物科学技术学院,甘肃兰州 730070;
2.甘肃肉羊繁育生物技术工程实验室,甘肃民勤 733300;
3.甘肃农业大学动物医学院,甘肃兰州 730070

收稿日期:2017-04-01 出版日期:2017-10-20 发布日期:2017-12-05
通讯作者: 马友记,E-mail:yjma@gsau.edu.cn
作者简介:高建锋(1991-),女,陕西榆林人,硕士研究生,从事动物遗传育种与繁殖研究。E-mail: 1518867937@qq.com
基金资助:
甘肃省科技重大专项计划(1602NKDH020); 甘肃农业大学校级课题(GSAU-ZL-2015-030); 国家肉羊产业技术体系项目(CARS-38)

Cloning, sequence analysis and tissue expression of MUSTN1 gene in sheep (Ovis aires)

GAO Jianfeng^{1, 2}, LU Zengkui^{1, 2}, MA Youji^{1, 2, *}, LI Taotao1^{1, 2}, ZHAO Xingxu³

1.College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
2.Gansu Engineering Laboratory of Sheep Breeding Biotechnology, Minqin 733300, China;
3.College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China

Received:2017-04-01 Online:2017-10-20 Published:2017-12-05

摘要/Abstract

摘要： 为了探讨绵羊(Ovis aires)MUSTN1基因特征及其在不同发育阶段的背最长肌与股二头肌组织中的表达规律,选取0、2、6、12和24月龄健康绵羊15只,每组3只,采取背最长肌和股二头肌组织,应用分子生物学等方法,对MUSTN1基因CDS区进行克隆,对其进行生物信息学和组织表达分析。结果表明,MUSTN1基因CDS区全序列为248 bp,编码82个氨基酸,其分子量为8.89 ku,等电点为9.93,GC含量大于AT含量,含有5个磷酸化位点、5个N-糖基化位点、1个O-糖基化位点、0个跨膜结构、疏水性蛋白,二级结构主要以无规卷曲为主;与山羊(Capra hircus)该基因核苷酸以及编码氨基酸序列的同源性最高,分别为99.2%、100%,其次为牛(Bos taurus),分别为97.2%、100%。MUSTN1基因在绵羊不同发育阶段(0、2、6、12和24月龄)的背最长肌与股二头肌中均有表达,在背最长肌中,24月龄MUSTN1表达量最高,极显著高于其他月龄(P<0.01),0月龄MUSTN1表达量最低,极显著低于2、12月龄(P<0.01),显著低于6月龄(P<0.05);而在6月龄绵羊股二头肌中MUSTN1表达量最高且极显著高于其他月龄(P<0.01),24月龄表达量次之且极显著高于12月龄(P<0.01),显著高于0月龄(P<0.05)。结论为绵羊MUSTN1基因的编码区序列在物种间具有保守性,且在不同发育阶段的背最长肌和股二头肌组织中均表达,其表达量的高低可能与骨骼肌肉的生长快慢相关。

关键词: MUSTN1, 绵羊, 克隆, 组织表达, 序列分析

Abstract: In order to study MUSTN1 gene characteristics of sheep (Ovis aires) and its expression at different developmental stages in longissimus dorsi and biceps femoris, fifteen sheep longissimus dorsi and biceps femoris tissues were collected at 0, 2, 6, 12 and 24 month old stages (three replicates for each). With application of molecular biology and other methods, the bioinformatics and tissue expression of the coding region of MUSTN1 gene were cloned and analyzed. The results showed that the CDS region of the MUSTN1 gene was 248 bp, encoded with 82 amino acid, molecular weight of 8.89 ku, its theoretical isoelectric point is 9.93, the GC content is greater than AT content, 5 phosphorylation sites, 5 N-glycosylation site, 1 O-glycosylation site, 0 transmembrane structure, hydrophobic protein and the secondary structure of MUSTN1 protein is dominated by random coil; the highest homology was 99.2% and 100% in the nucleotide and the amino acid sequence of goat (Capra hircus), followed by Bos taurus(97.2% and 100%, respectively). MUSTN1 gene was expressed in the longissimus dorsi and biceps femoris at different developmental stages (0, 2, 6, 12 and 24 month old). The expression of MUSTN1 in the longissimus dorsi was the highest at 24 month-old, which was significantly higher than those at other month old (P<0.01). The expression of MUSTN1 was the lowest at 0 month and significantly lower than that in 2 and 12 month old (P<0.01), significantly lower than that in 6 month old (P<0.05). The expression of MUSTN1 in 6 month old sheep biceps femoris was the highest and significantly higher than other month old (P<0.01). The expression level of 24 month old was significantly higher than that in 12 month old (P<0.01) and 0 month old (P<0.05). The conclusion was that the coding sequence of the MUSTN1 gene of sheep was conserved among species and expressed in the sheep of the dorsal long muscle and biceps femoris muscle at different development stages. The expression level of MUSTN1 gene may be related to the growth of skeletal muscle.

Key words: MUSTN1, sheep (Ovis aries), cloning, tissue expression, sequence analysis

中图分类号:

S826

高建锋, 卢曾奎, 马友记, 李讨讨, 赵兴绪. 绵羊MUSTN1基因的克隆、序列分析及其组织表达[J]. 浙江农业学报, 2017, 29(10): 1661-1668.

GAO Jianfeng, LU Zengkui, MA Youji, LI Taotao1, ZHAO Xingxu. Cloning, sequence analysis and tissue expression of MUSTN1 gene in sheep (Ovis aires)[J]. , 2017, 29(10): 1661-1668.

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绵羊MUSTN1基因的克隆、序列分析及其组织表达

Cloning, sequence analysis and tissue expression of MUSTN1 gene in sheep (Ovis aires)

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