静原鸡DERA基因生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2023.05.004

摘要/Abstract

摘要： 【目的】分析2-脱氧-D-核糖-5-磷酸醛缩酶(2-deoxyriboaldolase,DERA)CDS区序列特征及其在静原鸡不同组织和不同生长发育时期的表达水平,为进一步研究DERA基因在静原鸡肌肉中的调控机制提供参考。【方法】以42日龄静原鸡胸肌cDNA为模板,克隆DERA基因完整CDS区,测序并进行生物信息学分析。利用实时荧光定量PCR技术检测DERA基因在静原鸡不同生长发育阶段(7、42、126、180日龄)胸肌、腿肌和42日龄不同组织(心脏、脾脏、肝脏、肾脏、肺脏、胸肌和腿肌)中的表达水平。【结果】静原鸡DERA基因CDS区全长699 bp,编码232个氨基酸,与环颈雉的相似性最高,为99.1%,表明DERA蛋白氨基酸序列在进化过程中保守性较高。DERA属于稳定蛋白,平均亲水系数为0.175,属于疏水性蛋白;存在跨膜结构,没有信号肽,属于跨膜蛋白。蛋白结构域是一种deoC/LacD家族醛缩酶域,二级结构包含α-螺旋、延伸链和无规则卷曲。蛋白互作网络分析表明,DERA蛋白与PGM2、TPI1、TKT等蛋白存在互作关系。DERA基因在静原鸡肾脏、胸肌、腿肌、心脏、肝脏、肺脏、脾脏中均有表达,其中在肾脏和胸肌中的表达量较高,均显著高于其他组织(P＜0.05);在肺脏和脾脏中的表达量较低,均显著低于其他组织(P＜0.05);DERA基因在7日龄胸肌和腿肌组织中的表达量均显著高于42、126和180日龄(P＜0.05)。【结论】试验成功获得静原鸡DERA基因完整CDS区序列,该基因在静原鸡不同生长发育阶段胸肌和腿肌中的表达量以7日龄最高,且在42日龄静原鸡不同组织中以肾脏和胸肌中的表达量较高。结果为进一步研究DERA基因在静原鸡肌肉发育中的作用机制提供参考。

关键词: 静原鸡; DERA基因; 生物信息学; 表达

Abstract: 【Objective】 The aim of this study was to analyze the sequence characteristics of 2-deoxyriboaldolase (DERA) CDS region and its expression levels in different tissues and different growth stages,so as to lay a foundation for further study of the regulatory mechanism of DERA gene in the muscle of Jingyuan chickens.【Method】 The complete CDS region of DERA gene was cloned,sequenced and bioinformatics analyzed using the breast muscle cDNA of 42 days of age chickens as template.Real-time quantitative PCR was used to detect the expression of DERA gene in chest muscle,leg muscle and different tissues (heart,spleen,liver,kidney,lung,chest muscle and leg muscle) of Jingyuan chickens at different growth and development stages(7,42,126 and 180 days of age).【Result】 The CDS region of DERA gene in Jingyuan chickens was 699 bp in length,encoding 232 amino acids,and had the highest similarity (99.1%) with Phasianus colch.The results showed that the amino acid sequence of DERA protein was highly conserved during the evolutionary process.DERA was a stable protein,the average hydrophilic coefficient was 0.175,which was a hydrophobic protein.DERA protein had a transmembrane structure without signal peptide,which was a transmembrane protein.The protein domain was a deoC/LacD family aldolase domain.There were alpha helix,extended chain and random coil in secondary structure of DERA protein. Protein interaction network analysis showed that DERA protein interacted with PGM2,TPI1,TKT and other proteins.DERA gene was expressed in kidney,chest muscle,leg muscle,heart,liver,lung and spleen,among which the expression of kidney and chest muscle were higher,which were significantly higher than that of other tissues (P＜0.05),and the expression of lung and spleen were lower,which significantly lower than that of other tissues (P＜0.05).The expression of DERA gene in chest muscle and leg muscle at 7 days of age were significantly higher than that at 42,126 and 180 days of age (P＜0.05).【Conclusion】 The complete CDS sequence of DERA gene in Jingyuan chickens was successfully obtained in this experiment.The expression of DERA gene in chest muscle and leg muscle of Jingyuan chickens at different stages of growth and development was the highest at 7 days of age,and the expression of DERA gene in kidney and chast muscle of Jingyuan chickens were higher at 42 days of age.The results provided a reference for further study on the mechanism of DERA gene in muscle development of Jingyuan chickens.

Key words: Jingyuan chickens; DERA gene; bioinformatics; expression

中图分类号:

王浩睿, 禹保军, 杨嘉翊, 魏榕, 付希, 王川川, 张迪, 李德生, 蔡正云, 顾亚玲, 张娟. 静原鸡DERA基因生物信息学分析及组织表达研究[J]. 中国畜牧兽医, 2023, 50(5): 1764-1773.

WANG Haorui, YU Baojun, YANG Jiayi, WEI Rong, FU Xi, WANG Chuanchuan, ZHANG Di, LI Desheng, CAI Zhengyun, GU Yaling, ZHANG Juan. Bioinformatics Analysis and Tissue Expression of DERA Gene in Jingyuan Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1764-1773.

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