牦牛CAV-3基因的克隆及其在牦牛和黄牛组织的表达分析

Cloning of CAV-3 in Yak and Analysis Its Expression in Yak and Cattle

旨在对牦牛CAV-3基因进行克隆、生物信息学分析,并对其在牦牛组织中的表达规律进行初步研究。根据Gen Bank数据库中已知的黄牛CAV-3基因的m RNA序列并设计特异性引物,应用RT-PCR技术克隆牦牛CAV-3基因的编码区。运用生物信息学方法,分析并预测牦牛Caveolin-3蛋白的理化性质、疏水性、蛋白结构域以及蛋白质二级结构。通过半定量PCR技术检测CAV-3基因m RNA在牦牛和黄牛各组织中的表达;利用实时荧光定量PCR技术检测牦牛和黄牛肌肉组织中CAV-3基因m RNA表达水平。牦牛CAV-3的编码区全长631 bp,共编码151个氨基酸。CAV-3在牦牛肺、脾脏、肾脏、肝脏、卵巢组织中均不表达,仅在心脏和肌肉组织中表达,且在心脏组织的表达水平高于肌肉组织,CAV-3基因在黄牛各组织中的表达结果与牦牛一致。CAV-3基因在牦牛肌肉中的表达低于黄牛肌肉组织,但差异不显著(P>0.05)。

This study aims to clone, analyze bioinformatics and determine the expression pattern of CAV-3 gene in yak. A pair of special primers were designed according to released mRNA sequence of bovine CAV-3 in GenBank. A coding region sequence of yak CAV-3 was amplified by RT-PCR; the general physical and chemical properties, hydrophobicity, protein domains and protein secondary structures were systemically analyzed and predicted by bioinformatics techniques. The expression levels of CAV-3 mRNA in some organs of yak and cattle were detected by semi-quantitative PCR. Real-time PCR was employed to examine the expression levels of CAV-3 mRNA in yak and cattle muscles. The coding region sequence of CAV-3 gene in yak contains a complete ORF （ 631 bp ） which encoded 151 amino acids. CAV-3 mRNA expression in the heart and muscle was detected, but not in any other examined tissues, and its expression level in the heart was higher than in the muscle. The result in yak was consistent with that in cattle. Expression of CAV-3 gene in the yak muscle was less than in the cattle muscle, but not significantly （ P〉0.05 ） . The cloning and analysis of CAV-3 provided scientific basis for further study the function of CAV-3 gene in muscle development and muscle physiological process in the future.