鸭ARHGDIB基因克隆、生物信息学及组织表达研究

doi:10.16431/j.cnki.1671-7236.2025.07.004

摘要/Abstract

摘要： 【目的】旨在研究鸭Rho GDP解离抑制因子β(ARHGDIB)基因功能，进一步解析鸭皮下脂肪沉积的机理。【方法】以28日龄武禽10肉鸭肝脏组织cDNA为模板克隆ARHGDIB基因，经琼脂糖凝胶电泳及测序验证正确后，利用在线软件对所获序列进行比对及生物信息学分析，利用实时荧光定量 PCR检测ARHGDIB基因在鸭胸腺、肺脏、脾脏、心脏、十二指肠、肝脏、肾脏、腹脂、胸肌、腿肌、皮脂中的相对表达量。【结果】鸭ARHGDIB基因全长1 409 bp，预测其编码区长693 bp，包含7个外显子，可编码230个氨基酸，其中亮氨酸含量最高，占比为10%。鸭ARHGDIB基因核苷酸序列与鹅、牛、犬、山羊、斑马鱼、马、家猫、鸡、人、猕猴、家鼠、绵羊、黑猩猩、狒狒、野猪、野鸽的相似性分别为82.56%、50.53%、58.75%、54.08%、24.94%、56.30%、35.31%、81.79%、54.10%、54.90%、49.44%、47.25%、56.22%、55.99%、55.17%和82.64%。系统进化树分析表明，鸭与鹅的亲缘关系最近，其次为野鸽。鸭ARHGDIB蛋白属于稳定的亲水性蛋白，无跨膜结构，有24个磷酸化位点、2个N-糖基化位点，亚细胞定位预测蛋白主要定位在线粒体。鸭ARHGDIB蛋白二级结构由α-螺旋、延长链、β-转角和无规则卷曲4种类型组成，占比分别为25.22%、23.91%、4.35%和46.52%。三级结构与二级结构预测结果一致。实时荧光定量PCR结果显示，ARHGDIB基因在鸭各组织中均有表达，其中在胸腺中的表达量最高，且显著高于其他组织(P＜0.05)；其次是脾脏、心脏、肺脏；在皮脂中的表达量最低，且显著低于其他组织(P＜0.05)。【结论】鸭ARHGDIB基因编码区长693 bp，编码230个氨基酸，为亲水性蛋白，主要定位于线粒体。ARHGDIB基因在鸭各组织中均有表达，且在胸腺中表达量最高，皮脂中表达量最低。该研究为进一步探索鸭ARHGDIB基因的功能提供了理论依据，为解析其在脂肪代谢中的调节作用和鸭定向育种奠定基础。

关键词: 鸭; ARHGDIB 基因; 克隆; 生物信息学分析; 组织表达

Abstract: 【Objective】 This study aimed to investigate the functional role of the duck Rho GDP dissociation inhibitor β (ARHGDIB) gene and further elucidate the molecular mechanisms underlying subcutaneous fat deposition in ducks. 【Method】 The ARHGDIB gene was cloned from the liver tissue cDNA of 28-day-old Wuqin-10 meat ducks,and was verified to be correct by agarose gel electrophoresis and sequencing.The obtained sequences were compared and bioinformatics analyzed using online software.The relative expression of ARHGDIB gene in thymus,lung,spleen,heart,duodenum,liver,kidney,abdominal fat,pectoralis muscles,leg muscle,and subcutaneous fat of ducks was detected by Real-time quantitative PCR. 【Result】 The full length of ARHGDIB gene in ducks was 1 409 bp,with a predicted coding region of 693 bp spanning 7 exons,encoding 230 amino acids.Leucine was the most abundant amino acid (10% of the total composition).Nucleotide sequence similarity of the ARHGDIB gene in ducks with that of Anser cygnoides,Bos taurus,Canis lupus familiaris,Capra hircus,Danio rerio,Equus caballus,Felis catust,Gallus gallus,Homo sapiens,Macaca fascicularis,Mus musculus,Ovis aries,Pan troglodytes,Papio anubis,Sus scrofa,Columba livia were 82.56%,50.53%,58.75%,54.08%,24.94%,56.30%,35.31%,81.79%,54.10%,54.90%,49.44%,47.25%,56.22%,55.99%,55.17%,and 82.64%,respectively.The phylogenetic tree analysis showed that ducks were most closely related to Anser cygnoides,followed by Columba livia.The ARHGDIB protein in ducks was a stable hydrophilic protein lacking transmembrane domains.It contained 24 phosphorylation sites and 2 N-glycosylation sites.Subcellular localization prediction suggested that the protein was predominantly localized to the mitochondria.The secondary structure of the ARHGDIB protein in ducks comprises α-helices,extended chain,β-turns,and random coils,with respective proportions of 25.22%,23.91%,4.35% and 46.52%.The predicted tertiary structure aligned with the secondary structure.Real-time quantitative PCR results demonstrated that the ARHGDIB gene was ubiquitously expressed across all examined tissues of ducks,with the highest expression observed in the thymus,which was significantly higher than in other tissues (P＜0.05).Moderately high expression was detected in the spleen,heart,and lung,while subcutaneous fat showed the lowest expression,significantly lower than all other tissues (P＜0.05). 【Conclusion】 The coding region of ARHGDIB gene in ducks was 693 bp long and encodes 230 amino acids.ARHGDIB protein in ducks was hydrophilic and predominantly localized to mitochondria.The ARHGDIB gene was expressed in various tissues of ducks,with the highest levels in thymus and the lowest level in subcutaneous fat.These findings provided a theoretical foundation for further investigation of ARHGDIB gene function in ducks,and establish a basis for elucidating its regulatory role in lipid metabolism and supporting targeted duck breeding programs.

Key words: ducks; ARHGDIB gene; cloning; bioinformatics analysis; tissue expression

中图分类号:

S834

郑好, 刘梦云, 王明玉, 董霞, 史良玉, 余博, 杨宇, 周傲, 陈星. 鸭ARHGDIB基因克隆、生物信息学及组织表达研究[J]. 中国畜牧兽医, 2025, 52(7): 3004-3015.

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.

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