昆明犬IGF2基因克隆、生物信息学分析及其时空表达研究

doi:10.16431/j.cnki.1671-7236.2022.03.008

Abstract

Abstract: 【Objective】 This study was aimed to clone insulin-like growth factor 2 (IGF2) gene of Kunming dogs and obtain its sequence characteristics and spatio-temporal expression, to provide basic data for the study of body size and growth characteristics of working dogs.【Method】 The IGF2 gene CDS region of Kunming dogs was amplified by PCR technique, the structure and function of IGF2 protein in Kunming dogs were analyzed and predicted by bioinformatics method.The expression levels of IGF2 gene in heart, liver, spleen, lung, kindey and medial thigh muscle and liver at different growth stage in Kunming dogs were detected by Real-time quantitative PCR.【Result】 The CDS region of IGF2 gene in Kunming dogs was 717 bp which encoded 238 amino acids.The genetic tree analysis showed that the genetic distance of IGF2 gene from Kunming was closest to that of Red fox, and the genetic distance was farthest from that of chicken.The theoretical molecular weight and isoelectric point of IGF2 protein was 26.46 ku and 9.61, respectively.It had no signal peptide and transmembrane structure and belonged to hydrophilic non-secretory protein.The results of subcellular localization showed that IGF2 protein was mainly located in the nucleus (47.8%) and mitochondria (17.4%), and had 23 phosphorylation sites.The secondary structure of the protein was mainly composed of alpha helix and random coil.The prediction of tertiary structure was consistent with the results of secondary structure.The mRNA expression level of IGF2 gene in liver of 2.5 months old Kunming dogs was the highest, and was extremely significantly higher than that in kidney, lung and muscle (P<0.01).The expression of IGF2 gene in the liver of puppies (2.5-month-old) was extremely significantly higher than that of puppies (6-month-old), youth (1-year-old) and adults (1.7 and 2.5-year-old) (P<0.01).【Conclusion】 Kunming dog IGF2 gene was successfully cloned, and it was revealed that IGF2 was widely expressed in many tissues, which was highest in liver.The results could provide a basis for further study on the regulation mechanism of IGF2 gene in the growth and development of Kunming dogs.

Key words: insulin-like growth factor 2 (IGF2); Kunming dogs; cloning; sequence analysis; tissue expression

CLC Number:

WEI Yunfang, LI Feixiang, Xing Yun, Huang Qingguo, LI Judong, WAN Jiusheng, CHEN Fangliang. Study on Cloning, Bioinformatics Analysis and Spatio-temporal Expression of IGF2 Gene in Kunming Dogs[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 866-875.

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Study on Cloning, Bioinformatics Analysis and Spatio-temporal Expression of IGF2 Gene in Kunming Dogs

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