南方农业学报2024，Vol.55Issue(4)：1181-1193,13.DOI:10.3969/j.issn.2095-1191.2024.04.026
中华鳖IGF2和IGF2R基因克隆及其功能研究
Cloning and functional analysis of IGF2 and IGF2R genes in Chinese soft-shelled turtle(Pelodiscus sinensis)
摘要
Abstract
[Objective]To explore the role of insulin-like growth factor 2 gene(IGF2)and insulin-like growth factor 2 receptor gene(IGF2R)in the growth of Pelodiscus sinensis,and to lay a foundation for revealing the mechanism of IGFs system in the allometric growth and sexual growth difference of P.sinensis.[Method]The cDNA sequences of IGF2 and IGF2R genes in P.sinensis were cloned by RACE.Bioinformatics analysis was performed using online softwares such as BioEdit,ExPASy,TMHMM Server v2.0,SignalP and SMART.The expression changes of IGF2 and IGF2R genes in dif-ferent tissues,different phenotypes and different sex steroid hormones of male and female P.sinensis were analyzed by real-time fluorescence quantitative PCR.[Result]The total length of the cDNA sequence of IGF2 gene was 1180 bp,en-coding 222 amino acid residues.IGF2 protein,with a relative molecular weight of 24.84 kD and theoretical isoelectric point(pI)of 9.00,was a transmembrane protein and contained one IIGF domain.The total length of the cDNA sequence of IGF2R gene was 8765 bp,encoding 2443 amino acid residues.IGF2R protein had a relative molecular weight of 271.92 kD and a pI of 5.64.It was a transmembrane protein with fourteen repeated CIMR domains and one FN2 domain.Phylogenetic trees based on IGF2 and IGF2R amino acid sequence similarity showed that P.sinensis was most closely related to tortoises and far from fish and amphibians.IGF2 and IGF2R genes had a wide tissue expression profile in P.si-nensis.The relative expression of the IGF2 gene in testis was significantly higher than that in the ovary(P<0.05,the same below),while the situation was the opposite for IGF2R gene.The relative expression of IGF2 gene in the liver of fast growing P.sinensis individuals was significantly higher than that in the slow-growing individuals,while the expres-sion pattern of IGF2R gene was the opposite.Under estradiol(E2)treatment,except that the relative expression of IGF2R gene in the liver of female P.sinensis was significantly increased at 6 h after injection,the relative expression of IGF2 and IGF2R genes in the liver of male and female P.sinensis were decreased on the whole.MT treatment had no significant effect on IGF2 gene expression in male P.sinensis liver(P>0.05),but significantly increased the relative expression of IGF2 gene in female P.sinensis liver 48 h after injection,and significantly decreased the relative expression of IGF2R gene in female P.sinensis liver.[Conclusion]IGF2 and IGF2R genes are involved in the regulation of the gonad develop-ment and maturation of P.sinensis,and IGF2R may negatively regulate the growth of P.sinensis by internalizing and de-grading IGF2.In addition,IGF2 and IGF2R genes regulate the growth of P.sinensis by down-regulating their expres-sion levels in response to exogenous hormone stimulation.关键词
中华鳖/IGF2基因/IGF2R基因/生长差异/性类固醇激素Key words
Pelodiscus sinensis/IGF2 gene/IGF2R gene/growth difference/sex steroid hormone分类
农业科技引用本文复制引用
曾丹,李林,严峰,陈欣成,吴成瑶,李菁菁,涂洋洋,王晓清..中华鳖IGF2和IGF2R基因克隆及其功能研究[J].南方农业学报,2024,55(4):1181-1193,13.基金项目
国家自然科学基金项目(31672640) （31672640）
湖南省创新平台与人才计划科技特派员服务乡村振兴项目(2022NK4150) （2022NK4150）
湖南省教育厅科学研究项目(22C0385) （22C0385）
湖南文理学院博士启动项目(21BSQD12) National Natural Science Foundation of China(31672640) （21BSQD12）
Rural Science and Technology Spe-cial Correspodent Serving Rural Revitalization Project of Hunan Innovation Platform and Talent Plan(2022NK4150) （2022NK4150）
Science Research Project of Hunan Education Department(22C0385) （22C0385）
Doctoral Start-up Project of Hunan University of Arts and Sciences(21BSQD12) （21BSQD12）
