武定鸡GHRL基因克隆、组织表达及多态性分析

doi:10.16431/j.cnki.1671-7236.2025.11.026

摘要/Abstract

摘要： 【目的】克隆武定鸡胃饥饿素(ghrelin，GHRL)基因，阐明其组织表达差异及其功能性单核苷酸多态性(SNP)位点对生长性状的影响。【方法】克隆武定鸡GHRL基因编码区(CDS)并进行生物信息学分析，探究其分子结构和功能特性；利用实时荧光定量PCR检测GHRL基因在不同发育阶段(50、120和365日龄)不同组织中的表达差异，通过PCR产物直接测序技术检测GHRL基因内含子3、外显子4(652 bp)区域的SNP位点，分析其与武定鸡生长性状的关联性。【结果】成功克隆武定鸡GHRL基因CDS序列，其长度为351 bp，编码116个氨基酸。生物信息学分析显示，GHRL蛋白为亲水性不稳定蛋白，含有信号肽序列和跨膜结构域，且该蛋白包含Motilin_ghrelin和Motilin_assoc 2个保守结构域。实时荧光定量PCR结果显示，不同日龄武定鸡GHRL基因在腺胃中表达量显著高于其他组织(P＜0.05)，且120日龄时表达量显著高于50、365日龄(P＜0.05)。在GHRL基因内含子3中检测到3个SNPs位点(g.4374 C＞T、g.4375 C＞T、g.4431 G＞T)，在外显子4中检测到1个SNP(g.4629 G＞A)为异义替换。关联分析结果显示，g.4374 C＞T和g.4629 G＞A位点与武定鸡公鸡胫长及母鸡龙骨长和胸角显著相关(P＜0.05)。【结论】武定鸡GHRL基因在调节食欲和能量代谢、促进生长激素分泌和释放方面具有重要作用，其多态性与生长性状显著相关。研究结果为武定鸡的分子育种提供了重要理论依据。

关键词: 武定鸡; GHRL基因; 组织表达; 生长性状; 关联分析

Abstract: 【Objective】 The purpose of this experiment was to clone the ghrelin (GHRL) gene in Wuding chicken,clarify the differences in tissue expression and the effects of functional single nucleotide polymorphism (SNP) on growth traits. 【Method】 The coding sequence (CDS) of GHRL gene in Wuding chicken was cloned and subjected to bioinformatics analysis to reveal its molecular structure and functional properties.The expression differences of GHRL gene in different tissues at different developmental stages (50,120 and 365 days of age) were detected by Real-time quantitative PCR.SNP in the intron 3 and exon 4 region (652 bp) of GHRL gene were detected by direct sequencing technology,and their associations with growth traits of Wuding chicken were analyzed. 【Result】 The CDS sequence of GHRL gene in Wuding chicken was successfully cloned,with a length of 351 bp，encoding 116 amino acids residues.Bioinformatic characterization identified GHRL as a hydrophilic,unstable protein featuring both a signal peptide and transmembrane region,with two conserved domains (Motilin_ghrelin and Motilin_assoc).The results of Real-time quantitative PCR showed that the expression of GHRL gene in glandular stomach of Wuding chicken at different ages was significantly higher than that in other tissues (P＜0.05),and the expression at 120 days of age was significantly higher than that at 50 and 365 days of age (P＜0.05).Three SNPs (g.4374 C＞T,g.4375 C＞T and g.4431 G＞T) were detected in intron 3 of GHRL gene,and one missense substitution (g.4629 G＞A) was detected in exon 4.The association analysis results showed that g.4374 C＞T and g.4629 G＞A significantly correlated with the tibia length of roosters and the sternum length and breast angle of hens in Wuding chickens (P＜0.05). 【Conclusion】 GHRL gene in Wuding chicken played an important role in regulating appetite and energy metabolism,promoting the secretion and release of growth hormone,and was significantly correlated with growth traits.The results provided important theoretical basis for molecular breeding of Wuding chicken.

Key words: Wuding chicken; GHRL gene; tissue expression; growth traits; association analysis

中图分类号:

S831.2

周滟, 高瑞霞, 黄丽鸽, 范新阳, 保志鹏, 苗永旺. 武定鸡GHRL基因克隆、组织表达及多态性分析[J]. 中国畜牧兽医, 2025, 52(11): 5299-5309.

ZHOU Yan, GAO Ruixia, HUANG Lige, FAN Xinyang, BAO Zhipeng, MIAO Yongwang. Cloning,Tissue Expression,and Polymorphism Analysis of GHRL Gene in Wuding Chicken[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5299-5309.

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