绵羊CS-1基因的克隆及在凉山半细毛羊不同组织中的表达

Cloning of CS-1 in Ovine and Tissue Expression in Liangshan Semi-fine Sheep

旨在研究绵羊CS-1基因序列及其蛋白质的结构与功能以及其在不同组织中的表达情况,进而预测其与肉质性状的关联性。Calsarcins(Calcineurin-associated sarcomeric protein,CS)家族是一个与钙调神经磷酸酶(Calci-neurin,CaN)相结合的肌纤维特异表达蛋白新成员,CS-1基因是与畜禽肉质性状密切相关的重要候选基因。本研究根据牛、人和鼠CS-1基因mRNA,应用比较基因组学技术成功克隆了绵羊CS-1基因全长cDNA,对序列及编码的氨基酸进行生物信息学分析。结果显示,该基因cDNA全长951bp,完整的开放阅读框(ORF)为79～872bp,编码264个氨基酸。氨基酸序列中存在3个保守结构域,蛋白质二级结构以无规卷曲和α-螺旋为主,富含疏水区,存在多个磷酸化位点和1个蛋白激酶C(Protein kinase C,PKC)的磷酸化位点。通过实时荧光定量RT-PCR技术分析CS-1基因在凉山半细毛羊部分组织中的表达情况。结果表明:CS-1基因主要在凉山半细毛羊的心脏和背肌中表达,15d在心脏的表达量最高,极显著高于其它各组织(P<0.01);在心脏和背肌中,CS-1基因的表达量随着年龄的增长而减少,其中在60d时心脏中的表达量最高。研究结果为绵羊的肉质性状改善提供科学依据。

The aim of this study was to investigate CS-1 gene sequences,protein structure,function and its expression in different tissues,so as to predict its association with meat quality traits.Calsarcins（Calcineurin-associated sarcomeric protein,CS）is a family of calcium-binding proteins,CS-1 gene is closely related with meat quality in livestock and poultry.Based on the bovine,Mus musculus and Homo sapiens CS-1 mRNA sequences,the cDNA of CS-1 gene in ovine was amplified successfully by using comparative genomics technology.The results showed that CS-1 of ovine was 951 bp in length with an open reading frame（ORF）793 bp in length and encoded 264 amino acids,and there were three conserved domains in amino acids sequences.Bioinformation analysis indicated that its secondary structures mainly were random coil and helical regions,and contained rich hydrophobic regions,certain phosphorylation sites and protein kinase C（PKC） sites.Meanwhile,tissue expression of the CS-1 gene in the Semi-fine wool sheep were analyzed,the results showed that it was expressed in heart and muscle.When the Semi-fine Wool sheep was at fifteen days old,the expression was the highest in heart,and there was a significant difference between heart and other tissues（P0.01）.Furthermore,the expression of CS-1 decreased with the rise of the age and became the highest when the Semi-fine Wool sheep was at sixty days old in heart.This result could provide scientific basis for improve meat quality traits.