摘要：

旨在克隆甘南牦牛IGF-1基因及进行生物信息学分析并检测其在不同组织中的表达规律,为研究其功能提供参考。以甘南牦牛睾丸组织cDNA为模板,通过PCR技术扩增IGF-1基因CDS区,进而开展生物信息学分析,再运用实时荧光定量PCR技术检测其在心脏、肝脏、肺脏、肾脏、脾脏、肌肉、睾丸及脂肪组织中的表达规律。结果表明:甘南牦牛IGF-1基因CDS区全长567 bp,编码188个氨基酸;IGF-1蛋白分子式为C₉₀₇H₁₄₆₄N₂₇₂O₂₆₅S₁₇,等电点为 9.72,不稳定指数为69.91,属于不稳定蛋白;IGF-1蛋白为含有信号肽的亲水性分泌蛋白,主要定位于细胞外,且含有10个O-糖基化位点与19个磷酸化位点;二级结构以无规则卷曲和α-螺旋为主;与IGF1R、IGFBP3、INSR及IGFBP1等蛋白存在相互作用。实时荧光定量PCR结果显示,IGF-1基因在甘南牦牛肝脏中的表达量极显著高于其他组织,脾脏和脂肪中的表达量极显著高于肺脏、睾丸及肌肉。

关键词: 甘南牦牛, IGF-1基因, 克隆, 生物信息学, 组织表达

Abstract:

The aim was to clone the IGF-1 gene of Gannan yak,conduct bioinformatics analysis and detect its expression pattern in different tissues,so as to provide a reference for studying its function.Taking the cDNA of testicular tissue of Gannan yaks as the template,the CDS region of the IGF-1 gene was amplified by PCR,and then bioinformatics analysis was carried out.Subsequently,Quantitative Real-time PCR(RT-qPCR)was used to detect its expression patterns in heart,liver,lung,kidney,spleen,muscle,testicular and adipose tissues.The results showed that the CDS region of the IGF-1 gene was Gannan yak was 567 bp in total length and encoded 188 amino acids.The molecular formula of IGF-1 protein was C₉₀₇H₁₄₆₄N₂₇₂O₂₆₅S₁₇,with an isoelectric point of 9.72 and an instability index of 69.91,which belonged to unstable proteins.The IGF-1 protein was a hydrophilic secreted protein containing signal peptides.It was mainly located outside the cell and contained 10 O-glycosylation and 19 phosphorylation sites.The secondary structure was mainly composed of random coil and α-helix.It interacted with proteins such as IGF1R,IGFBP3,INSR and IGFBP1.The results of RT-qPCR showed that the expression level of IGF-1 gene in the liver of Gannan yak was extremely significantly higher than that in other tissues,and the expression levels in the spleen and fat were extremely significantly higher than those in the lungs,testicles and muscles.

Key words: Gannan yak, IGF-1 gene, Cloning, Bioinformatics, Tissue expression

中图分类号: 

• S823

• Q78基因工程(遗传工程)