绵羊CS-3基因的克隆及其在绵羊不同组织中的表达研究

Cloning of CS-3 Gene and its Expression in Different Tissues in Ovine

试验首次克隆了绵羊CS-3基因序列,预测了CS-3蛋白的结构、功能及其特点,并分析了其在不同组织中的表达情况及CS-3基因与肉质性状的相关性。结果显示,CS-3基因cDNA全长838 bp,完整的开放阅读框(ORF)为70-804 bp,编码244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技术分析了CS-3基因在凉山半细毛羊部分组织中的表达情况。结果表明,CS-3基因主要在凉山半细毛羊的心脏和肌肉中表达,15 d时各组织中心脏的表达量最高,极显著高于其它各组织(P<0.01);在心脏和肌肉组织中,CS-3基因的表达量随着年龄的增长而减少,其中在60 d时心脏中的表达量最高。

This study was to clone CS-3 gene for the first time with its expression in different tissues,and predict its protein structure and fuction,so as to analyze the meat quality traits associated with.The results showed that CS-3 of ovine was 838 bp in length with an open reading frame（ORF） 735 bp long and encoded 244 amino acids,and there were three conserved domains in amino acids sequences.Bioinformation analysis indicated 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 gene in Liangshan semi-fine wool sheep were analysed,results showed that it was expressed in heart and muscle.When the ovine was of fifteen-days age,the expression was the highest in heart,and there was very significantly different between heart and other internal organ tissues（P〈0.01）.Furthermore,the expression of CS-3 decreased with the rise of the age and became the highest when the ovine was at sixty-days age in heart.