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

Abstract

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

CLC Number:

S831.2

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.

References

[1] KOJIMA M，HOSODA H，DATE Y，et al.Ghrelin is a growth-hormone-releasing acylated peptide from stomach[J].Nature，1999，402(6762)：656-660.
[2] DATE Y，KOJIMA M，HOSODA H，et al.Ghrelin，a novel growth hormone-releasing acylated peptide，is synthesized in a distinct endocrine cell type in the gastrointestinal tracts of rats and humans[J].Endocrinology，2000，141(11)：4255-4261.
[3] GUALILLO O，LAGO F，GÓMEZ-REINO J，et al.Ghrelin，a widespread hormone：Insights into molecular and cellular regulation of its expression and mechanism of action[J].FEBS Letters，2003，552(2-3)：105-109.
[4] HOSODA H，KOJIMA M，MATSUO H，et al.Purification and characterization of rat des-Gln14-Ghreli，a second endogenous ligand for the growth hormone secretagogue receptor[J].The Journal of Biological Chemistry，2000，275(29)：21995-22000.
[5] DELHANTY P J，NEGGERS S J，VAN DER LELY A J.Mechanisms in endocrinology：Ghrelin：The differences between acyl- and des-acyl ghrelin[J].European Journal of Endocrinology，2012，167(5)：601-608.
[6] GUAN X M，YU H，PALYHA O C，et al.Distribution of mRNA encoding the growth hormone secretagogue receptor in brain and peripheral tissues[J].Molecular Brain Research，1997，48(1)：23-29.
[7] DATE Y，MURAKAMI N，TOSHINAI K，et al.The role of the gastric afferent vagal nerve in ghrelin-induced feeding and growth hormone secretion in rats[J].Gastroenterology，2002，123(4)：1120-1128.
[8] LIM C T，KOLA B，KORBONITS M.The ghrelin/GOAT/GHS-R system and energy metabolism[J].Reviews in Endocrine & Metabolic Disorders，2011，12(3)：173-186.
[9] FANG M，NIE Q，LUO C，et al.An 8 bp indel in exon 1 of Ghrelin gene associated with chicken growth[J].Domestic Animal Endocrinology，2007，32(3):216-225.
[10] 何丹林，方梅霞，聂庆华，等.鸡Ghrelin基因C2100T位点与生长和脂肪性状的相关性[J].广东农业科学，2007，4：73-75.HE D L,FANG M X,NIE Q H,et al.Association of Ghrelin gene C2100T polymorphism with chicken growth and fat traits[J].Guangdong Agricultural Sciences，2007，4：73-75.(in Chinese)
[11] 李俊营，詹凯，许月英，等.巢湖鸭Ghrelin基因外显子3的单核苷酸多态性及其对屠体性状的影响[J].安徽农业科学，2010，38(14):7379-7381.LI J Y，ZHAN K，XU Y Y，et al.Effects of SNP in the exon 3 of Ghrelin gene on carcass traits in Chaohu duck[J].Journal of Anhui Agricultural Sciences，2010，38(14)：7379-7381.(in Chinese)
[12] 王宗礼.畜禽遗传资源调查技术手册[M].北京：中国农业出版社，2019.WANG Z L.Technical Manual for Investigation of Livestock and Poultry Genetic Resources[M].Beijing：China Agriculture Press，2019.(in Chinese)
[13] 中华人民共和国农业农村部.家禽生产性能名词术语和度量计算方法：NY/T 823—2020[S].北京：中国农业出版社，2020.MINISTRY OF AGRICULTURE AND RURAL AFFAIRS OF THE PEOPLE’S REPUBLIC OF CHINA.Poultry production performance terminology and measurement calculation method：NY/T 823—2020[S].Beijing：China Agriculture Press，2020.(in Chinese)
[14] ITOH Z.Motilin and clinical application[J].Peptides，1997，18(4)：593-608.
[15] KOJIMA M，HOSODA H，MATSUO H，et al.Ghrelin：Discovery of the natural endogenous ligand for the growth hormone secretagogue receptor[J].Trends in Endocrinology & Metabolism，2001，12(3)：118-122.
[16] LIM C T，KOLA B，GROSSMAN A，et al.The expression of ghrelin O-acyltransferase (GOAT) in human tissues[J].Endocrine Journal，2011，58(8)：707-710.
[17] BRIGGS D I，ENRIORI P J，LEMUS M B，et al.Diet-induced obesity causes ghrelin resistance in arcuate NPY/AgRP neurons[J].Endocrinology，2010，151(10)：4745-4755.
[18] HOWARD A D，FEIGHNER S D，CULLY DF，et al.A receptor in pituitary and hypothalamus that functions in growth hormone release[J].Science，1996，273(5277)：974-977.
[19] SEOANE L M，TOVAR S，BALDELLI R，et al.Ghrelin elicits a marked stimulatory effect on GH secretion in freely-moving rats[J].European Journal of Endocrinology，2000，143(5)：R7-R9.
[20] WREN A M，SMALL C J，ABBOTT C R，et al.Ghrelin causes hyperphagia and obesity in rats[J].Diabetes，2001，50(11)：2540-2547.
[21] KOJIMA M，KANGAWA K.Ghrelin：Structure and function[J].Physiological Reviews，2005，85(2)：495-522.
[22] WANG W，CHENG L，GUO J，et al.Expression of Ghrelin in gastrointestinal tract and the effect of early weaning on Ghrelin expression in lambs[J].Molecular Biology Reports，2014，41(2)：909-914.
[23] CALO J，BLANCO A M，COMESAÑA S，et al.First evidence for the presence of amino acid sensing mechanisms in the fish gastrointestinal tract[J].Scientific Reports，2021，11(1)：4933.

Cloning,Tissue Expression,and Polymorphism Analysis of GHRL Gene in Wuding Chicken

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	XU Yuanyuan, XIE Yingxue, LU Xingrong, FENG Chao, SHANG Jianghua. Sequence Analysis, Overexpression Vector Construction and Tissue Expression of BTG2 Gene in Buffalo [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4009-4020.
[2]	ZHANG Haijuan, ZHAO Yong, YANG Hongwen, WU Yan, XIANG Jin, FU Hongmei, DING Yijie, ZHANG Yiyu. Polymorphism of GH1 Gene and Its Association with Growth Traits and Functional Prediction of GH1 Protein in Guanling Cattle [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4226-4236.
[3]	LI Zhiquan, GAO Mengruo, WEN Yijun, FU Sijing, XU Shuping, YANG Taotao, ZHANG Zhibang, FANG Xia, LI Kai, LI Pengcheng. Cloning,Bioinformatics Comparative Analysis and Tissue Expression of HSPB1 Gene Between Wuyi Black Pig and Duroc Pig [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4248-4259.
[4]	ZHU Shuo, SUN Jinghua, BAO Jingjing, SHANG Mingyu, LIU Tianyi, XIONG Jinke, YANG Peifu, YAO Shuhai, SUN Xintian, HE Jianning, YANG Huiguo, ZHANG Li. Polymorphisms of MAP2K6 Gene and Its Association Analysis with Growth Traits in Hu Sheep [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3695-3706.
[5]	ZHENG Hao, LIU Mengyun, WANG Mingyu, DONG Xia, SHI Liangyu, YU Bo, YANG Yu, ZHOU Ao, CHEN Xing. Cloning, Bioinformatics and Tissue Expression Analysis of ARHGDIB Genes in Ducks [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3004-3015.
[6]	LI Zhiquan, FU Sijing, XU Shuping, GAO Mengruo, YANG Taotao, ZHANG Zhibang, LI Kai, LI Pengcheng. Cloning, Bioinformatics Analysis and Tissue Expression of HSPA5 Gene in Wuyi Black Pig [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3214-3224.
[7]	CHU Hongen, LIU Yuan, FENG Xue, BAI Xue, YANG Mengli, LI Juan, HE Lixia, LIU Shuang, FENG Lan, MA Yun. Cloning,Bioinformatics and Tissue Expression Analysis of Bovine TGFB1 Gene [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2506-2518.
[8]	LI Tianxiu, LI Xinpeng, DONG Xinxing, LAN Guoxiang, YAN Dawei, ZHU Jiawei. Amplification,Sequence Analysis and Tissue Expression Study of HMOX2 Gene in Lijiang Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2219-2231.
[9]	ZHAO Zhonglong, ZHANG Yong, YANG Hong, YANG Runqian, AI Zhaobi, YE Li, SHU Chang. Analysis of PPP3CA Gene Polymorphism and Its Association with Body Size Traits in Zhijin White Geese [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1729-1738.
[10]	AI Xiaonan, CHENG Lifen, BAI Pu, CHEN Zhenghao, XUE Lina, ZHOU Shenghua. Cloning and Bioinformatics Analysis of ACSL1 Gene and Its Expression Pattern During Adipocyte Differentiation in Lingqiu Qingbei Goats [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(2): 499-511.
[11]	YANG Jinlin, LI Hui, ZHAO Depeng, SHI Yushi, LONG Xia, TAN Qisong. Polymorphisms of EDNRB2 Gene and Its Association with Skin Color Traits in Chishui Black-bone Chickens [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(2): 759-770.
[12]	YAN Nana, GULIZGULINA Asghar, HANIKZ Tulav, SMUBATI Guli Shahatynur, GURSIYAR Kanish, MA Junde, WU Weiwei, LIU Lingling. Study on Genomic Selection of Growth and Wool Traits in Chinese Merino Sheep [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4776-4784.
[13]	XIE Beiyiting, WANG Yue, MENG Chunhua, QIAN Yong, ZHANG Jun, ZHANG Jianli, WANG Huili, CAO Shaoxian, LI Yinxia. Cloning and Sequence Analysis of PSMB9 Gene in Hu Sheep and Its Effect on Myoblast Proliferation [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(1): 1-12.
[14]	JIANG Lei, WANG Yuxin, WAN Yuan, ZAN Linsen, WANG Hongbao. Bioinformatics and Expression Characteristics Analysis of MGP Gene in Qinchuan Cattle [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(6): 2261-2272.
[15]	SUN Shiyu, LI Jinlan, XING Jiani, DONG Yaohui, LI Yanyan, WANG Youli, LIN Yaqiu. Cloning,Bioinformatics Analysis and Spatiotemporal Expression of CKMT2 Gene in Jianzhou Big-eared Goats [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(6): 2342-2353.