昆明犬IGF1R基因克隆及组织表达分析.

【Objective】 This study aimed to explore the IGF1R gene sequence structural characteristics and tissue expression in Kunming dogs, and to provide a basis for further study on the regulation mechanism of IGF1R gene in the growth and development and to lay a foundation for molecular genetics and breeding of Kunming dogs. 【Method】The IGF1R gene sequence of Kunming dogs was amplified and sequenced based on molecular cloning technology. Sequence homology alignment and phylogenetic tree of IGF1R gene were constructed by bioinformatics method. The physical and chemical property, sequence structural characteristics of the corresponding amino acid sequence were analyzed by online predicition software, and the tertiary structure model of IGF1R protein were predicted. Meanwhile, real-time quantitative PCR was used to detect the relative expression of IGF1R gene in various tissues of Kunming dogs. 【Result】The CDS region of IGF1R gene in Kunming dogs was 4104 bp encoding 1367 amino acids. IGF1R gene CDS in Kunming dogs had higher homology with Canis lupus familiaris(100%) and Vulpes vulpes(99.3%), but lower homology with Gallus gallus(78.8%). The molecular weight and theoretical isoelectric point of IGF1 R protein were 154 890.07 ku and 5.58, respectively.It contained three fibronectin type(FN3) domains, one furin like cysteine-rich(FU) domain and tyrosine kinase(TyrKc) domain, was a hydrophilic transmembrane protein. Subcellular localization analysis showed that IGF1 R protein was evenly distributed on endoplasmic reticulum, Golgi apparatus and plasma membrane, with the proportion of 33.3%, respectively. The analysis of phosphorylation sites showed that there were 123 phosphorylation sites in IGF1 R protein. The secondary structure of the protein was mainly composed of α-helix and random coil. Tissue expression showed that IGF1R gene was expressed in different tissues of Kunming dogs, especially in lung, heart and kidney. 【Conclusion】 IGF1R gene had strong conservatism in the process of biological evolution. IGF1 R protein of Kunming dogs had the characteristic domain of IGF receptor protein and belonged to hydrophilic transmembrane protein.IGF1 R gene was expressed in 6 tissues of Kunming dog, especially in heart, lung and kidney. [ABSTRACT FROM AUTHOR]