昆明犬IGF-1基因CDS区克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2020.06.003

Abstract

Abstract: This study was aimed to systematically analyze the gene structure and function of insulin-like growth factor-1 (IGF-1) in Kunming dogs,in order to better understand the gene tissue expression pattern,and provide basic data for the study of body size and growth characteristics of working dogs.Kunming dogs was taken as the research object to amplify the CDS region of IGF-1 gene by molecular cloning technique.Sequence homology alignment and phylogenetic tree of IGF-1 gene were constructed by bioinformatics method.The physical and chemical property,subcellular structure,hydrophilicity and hydrophobicity and phosphorylation site of the corresponding amino acid sequence were analyzed by online predicition software.The secondary and tertiary structures of IGF-1 protein were predicted by SOPMA and SWISS-MODEL software.Meanwhile,the expression of IGF-1 gene in different tissues of Kunming dogs was detected by Real-time quantitative PCR.The results showed that the CDS region of IGF-1 gene in Kunming dogs was 462 bp which encoding 153 amino acids.IGF-1 gene CDS in Kunming dogs had higher homology with Canis lupus familiaris (100%),but lower homology with Mus musculus and Gallus gallus (82.6% and 81.2%).The theoretical molecular weight and isoelectric point of IGF-1 protein was 16.97 ku and 9.36,respectively.It was a hydrophilic and secretory protein with no transmembrane structure and its signal peptide region was 1-48 amino acids.The results of subcellular localization showed that IGF-1 protein was mainly located in the nucleus (56.5%) and mitochondria (17.4%).There were 15 phosphorylation sites.The secondary structure of the protein was mainly composed of alpha helix and random coil.Tissue expression showed that IGF-1 gene was expressed in different tissues of Kunming dogs,especially in spleen,liver and kidney.The results laid a theoretical reference for a better study of IGF-1 gene functions,and provided certain basic data for further exploration of growth and development characters of IGF-1 gene in Kunming dogs.

Key words: Kunming dogs; IGF-1 gene; cloning; tissue expression; bioinformatics analysis

CLC Number:

WEI Yunfang, LI Feixiang, MA Weiguo, XU Tao, LI Judong, CHEN Fangliang, WAN Jiusheng, CHEN Chao. Cloning,Bioinformatics Analysis and Tissue Expression Detection of IGF-1 Gene CDS Regions in Kunming Dogs[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(6): 1659-1667.

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Cloning,Bioinformatics Analysis and Tissue Expression Detection of IGF-1 Gene CDS Regions in Kunming Dogs

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