陆川猪Myf5基因克隆及其在背最长肌中发育性变化表达研究
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摘要
为了获得广西陆川猪生肌调节因子(Myf5)基因编码区序列(CDS),并探讨该基因在陆川猪背最长肌中的发育性表达情况,试验以陆川猪背最长肌组织总RNA为模板,克隆获得Myf5基因CDS,运用生物信息学软件进行测序及分析,并利用实时荧光定量PCR技术检测30,150,180,240,300日龄陆川猪背最长肌中Myf5基因的相对表达量。结果表明:试验克隆得到的陆川猪Myf5基因CDS全长为768 bp;与GenBank上公布的香猪(KC456667.1)Myf5基因CDS的同源性为99.6%;陆川猪存在第205位点的C→A,导致亮氨酸突变为蛋氨酸;广西陆川猪与香猪的遗传距离最近,它们聚为同一支;陆川猪Myf5蛋白氨基酸组成中丝氨酸含量最高,占氨基酸总数的13.7%;Myf5蛋白具有较强的亲水性,其蛋白高级结构包含α-螺旋、延伸链和无规则卷曲;陆川猪背最长肌Myf5基因相对表达量表现为30日龄时最高,在30～240日龄呈下降趋势,240～300日龄呈上升趋势。
In this study, the total RNA of the longissimus dorsi muscle of Luchuan pig was used as a template to clone the Myf5 gene CDS,and bioinformatics software was used to sequence and analyze it.The relative expression of Myf5 gene in the longissimus dorsi muscle of 30, 150, 180, 240, 300 days old Luchuan pig were detected by Real-time PCR. The results show that the total length of Myf5 gene CDS of Luchuan pig cloning is 768 bp. The homology of Myf5 gene CDS of Luchuan pig and Xiang pigs published on GenBank(KC456667.1) is 99.6%. Luchuan pigs have a C → A mutation at point 205, resulting in a leucine mutation to methionine.The genetic distance between Guangxi Luchuan pigs and Xiang pigs is the closest.They are in the same branch.The content of serine of Myf5 protein in Luchuan pig is the highest, accounting for 13.7% of the total amino acids. Myf5 protein has strong hydrophilicity, and its advanced protein structure includes analpha helix, an extended chain, and a random coil.The relative expression of the Myf5 gene of the longissimus dorsi muscle of Luchuan pig is the highest at the age of 30 days, and it is declining at the age of 30 to 240 days, and is increasing at the age of 240 to 300 days.
In this study, the total RNA of the longissimus dorsi muscle of Luchuan pig was used as a template to clone the Myf5 gene CDS,and bioinformatics software was used to sequence and analyze it.The relative expression of Myf5 gene in the longissimus dorsi muscle of 30, 150, 180, 240, 300 days old Luchuan pig were detected by Real-time PCR. The results show that the total length of Myf5 gene CDS of Luchuan pig cloning is 768 bp. The homology of Myf5 gene CDS of Luchuan pig and Xiang pigs published on GenBank(KC456667.1) is 99.6%. Luchuan pigs have a C → A mutation at point 205, resulting in a leucine mutation to methionine.The genetic distance between Guangxi Luchuan pigs and Xiang pigs is the closest.They are in the same branch.The content of serine of Myf5 protein in Luchuan pig is the highest, accounting for 13.7% of the total amino acids. Myf5 protein has strong hydrophilicity, and its advanced protein structure includes analpha helix, an extended chain, and a random coil.The relative expression of the Myf5 gene of the longissimus dorsi muscle of Luchuan pig is the highest at the age of 30 days, and it is declining at the age of 30 to 240 days, and is increasing at the age of 240 to 300 days.
引文
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[12] 高玲,谷大海,徐志强,等.MRFs基因家族对肉品质影响的研究进展[J].黑龙江畜牧兽医,2015(05上):65-67.
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[14] 赵丹璐,卢珊,张静,等.山猪MYF5基因SNPs及其形成的单倍型对部分肉质性状的影响[J].中国畜牧杂志,2014,50(15):19-23.
[2] 曹婷,侯冠彧,施力光,等.五指山猪不同组织中Myf5与MyoD1基因的表达研究[J].家畜生态学报,2012,33(5):12-15.
[3] 袁恬,冉雪琴,王芳,等.三个猪品种Myf5基因的克隆及序列分析[J].山地农业生物学报,2013,32(2):114-118.
[4] 黎威,高鹏飞,石建中,等.Myf5基因在马身猪和大白猪背最长肌中的发育性表达研究[J].畜牧兽医学报,2014,45(12):2081-2087.
[5] 李琼华.陆川猪优良特性和杂交开发利用[J].养猪,2010(6):36.
[6] 张家富,何若钢,江永强,等.陆川猪胴体品质及肉质的研究[J].养猪,2010(6):33-36.
[7] 晁哲,王峰,魏立民,等.海南五指山猪Myf5基因克隆与序列分析[J].黑龙江畜牧兽医,2014(05上):71-73.
[8] 孙文浩,朱庆.生肌决定因子Myf5基因的研究进展[J].黑龙江畜牧兽医,2008(7):28-30.
[9] 丁国杰,刘娣,李景芬.猪Myf5基因外显子1的多态性分析[J].江苏农业科学,2006(3):125-127.
[10] 豆腾飞,佟荟全,荣华,等.大围山微型鸡和艾维茵肉鸡骨骼肌卫星细胞增殖分化及Myf5基因表达差异研究[J].中国家禽,2017,39(11):8-12.
[11] 孙文浩.鸡Myf5和Myf6基因SNP多态性及其与屠宰性状和肉质性状的相关性分析[D].雅安:四川农业大学,2008.
[12] 高玲,谷大海,徐志强,等.MRFs基因家族对肉品质影响的研究进展[J].黑龙江畜牧兽医,2015(05上):65-67.
[13] 王巧巧,王亚南,唐辉,等.鸡肌纤维发育相关基因研究进展[J].家禽科学,2018(5):43-48.
[14] 赵丹璐,卢珊,张静,等.山猪MYF5基因SNPs及其形成的单倍型对部分肉质性状的影响[J].中国畜牧杂志,2014,50(15):19-23.