撒坝猪MCUR1基因克隆、生物信息学分析及其组织表达量检测

Cloning, bioinformatics analysis and tissue expression detection of MCUR1 gene in Saba pig

本研究克隆了撒坝猪MCUR 1基因,对其进行生物信息学分析,并通过实时荧光定量PCR检测其在撒坝猪、大白猪不同组织中的表达水平。根据GenBank上公布的猪的MCUR1基因序列(登录号:XM_003128183.4)设计引物,通过PCR扩增及测序获得撒坝猪MCUR 1基因CDS序列,该序列全长1095 bp,编码364个氨基酸;MCUR1蛋白的分子式C1787H2949N533O503S14,相对分子质量40.398 ku,理论等电点(pI)10.27,不稳定系数55.70,脂肪指数95.99,平均亲水性为-0.213,属于亲水蛋白;MCUR1蛋白没有信号肽和跨膜结构,含有2个螺旋卷曲结构,主要在细胞质中发挥生物学功能;含有32个磷酸化位点;含有一个CpG island;二级结构由58.24%的α-螺旋,4.12%的β-转角,30.22%的无规则卷曲,7.42%的延伸链构成;猪MCUR 1基因编码区序列与牛、鸡、狗、山羊、人、猴、鼠、绵羊的同源性分别为85.3%、68.0%、83.6%、87.9%、84.4%、80.3%、76.8%、88.2%,与绵羊、山羊、牛的亲缘关系较近,与鸡的关系较远;在肝脏中的表达量最多,肺脏中的表达量最少。在大白猪的背最长肌中MCUR 1的表达量高于撒坝猪的背最长肌(P<0.01)。

The aim of this study was to clone the MCUR 1 gene from Saba pigs,analyze it by bioinformatics,and detect its expression level in different tissues of Saba pig and Large White pig by real-time fluorescent quantitative PCR.According to the MCUR 1 gene sequence published in GenBank(accession number:XM_003128183.4),the primers were designed,and the MCUR 1 gene CDS sequence of Saba pig was obtained by PCR amplification and sequencing.The sequence was 1095 bp in length and encoded 364 amino acids.The molecular formula of MCUR1 protein is C1787H2949N533O503S14.The molecular weight is 40.398 ku,the theoretical isoelectric point(pI)is 10.27,the instability coefficient is 55.70,the fat index is 95.99,and the average hydrophilicity is-0.213,which belongs to hydrophilic protein.MCUR1 protein has no signal peptide and transmembrane structure,and contains two spiral coiled structure that function primarily in the cytoplasm;contains 32 phosphorylation sites;contains one CpG island;the secondary structure consists of 58.24%alpha-helix,4.12%beta turn,30.22%random coil and 7.42%extended chain.The homology of the coding region of the porcine MCUR 1 gene with bovine,chicken,dog,goat,human,monkey,mouse,and sheep was 85.3%,68.0%,83.6%,87.9%,84.4%,80.3%,76.8%and 88.2%,respectively,showing close relationship with sheep,goat and cattle,and distant relationship with chicken.The highest expression level of MCUR 1 was observed in the liver and the least expression was in the lung.The expression of MCUR 1 in the longissimus dorsi of the Large White pig was higher than that of the longissimus dorsi of the Saba pig(P<0.01).