生物技术通报 ›› 2021, Vol. 37 ›› Issue (8): 203-212.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1410

• 研究报告 • 上一篇    下一篇

鸡CEBPA基因CDS区克隆、表达及生物信息学分析

杜振伟(), 朱帅鹏, 马向飞, 李东华(), 孙桂荣   

  1. 河南农业大学动物科技学院,郑州 540046
  • 收稿日期:2020-11-19 出版日期:2021-08-26 发布日期:2021-09-10
  • 作者简介:杜振伟,男,硕士,研究方向:动物遗传育种与繁殖;E-mail: x13592547117@126.com
  • 基金资助:
    河南省自然科学青年科学基金项目(20230041020);中原学者康相涛科学家工作室(30601985)

Cloning,Expression and Bioinformatics Analysis of the CDS Region of Chicken CEBPA Gene

DU Zhen-wei(), ZHU Shuai-peng, MA Xiang-fei, LI Dong-hua(), SUN Gui-rong   

  1. College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 540046
  • Received:2020-11-19 Published:2021-08-26 Online:2021-09-10

摘要:

为了研究增强子结合蛋白α(CCAAT/Enhancer Binding Protein Alpha,CEBPA)的氨基酸序列特征,以及其编码基因在固始鸡不同组织中的mRNA表达谱。采用PCR技术克隆得到CEBPA的CDS序列全长975 bp,可编码氨基酸325个。生物信息学分析结果显示CEBPA不存在跨膜结构和信号肽,蛋白结构主要由无规则卷曲和α-螺旋结构构成,且多分布在细胞核中;蛋白互做网络构建分析表明,该蛋白与主要与TRBs家族、Runx转录因子家族等相互作用;氨基酸序列对比发现,鸡和鹌鹑的亲缘关系较近;鸡CEBPA 5'调控区-2 000 bp序列上启动子和CpG岛预测发现,该序列上有13个不同的启动子和一个长度1 640 bp的CpG岛;qRT-PCR表明,CEBPA在12周龄固始鸡不同组织中胸肌和腿肌的表达量显著低于其他组织(P<0.05);不同发育时期胸肌中22周龄的表达量显著低于6周龄和55周龄的表达量(P<0.05);且CEBPA在罗斯鸡胸肌的表达量显著高于固始鸡胸肌的表达量(P<0.05)。以上结果为进一步深入探讨鸡CEBPA在胸肌的脂质沉积过程中的生物学功能奠定了基础。

关键词: 固始鸡, CEBPA, 生物信息学分析, 胸肌, 表达分析

Abstract:

This work is to investigate the amino acid sequence characteristics of CCAAT/Enhancer Binding Protein Alpha(CEBPA)and the mRNA expression profile of its coding gene in different tissues of Gushi chicken. The CDS sequence of CEBPA with a total length of 975 bp was cloned by PCR,and it encoded 325 amino acids. The bioinformatics analyses showed that there was no transmembrane structure and signal peptide in CEBPA,and the protein structure was mainly composed of irregular crimp and α-spiral,and most of them were distributed in the nucleus. The construction of protein interaction network indicated that CEBPA might interact with TRBs family and Runx transcription factors family. Amino acid sequence comparison found that chicken and quail were closely related and had 13 different promoters and a 1 640 bp CpG island on its -2 000 bp sequence of the 5' regulatory region. qRT-PCR results showed that the expression of CEBPA gene in the breast muscle and leg muscle was significantly lower than that in other tissues of 12-week Guishi chickens(P<0.05). The expression of CEBPA in the breast muscle of 22-week chicken was significantly lower than that in 6-week and 55-week ones(P < 0.05). The expression of CEBPA in the breast muscle of Ross chicken was always significantly higher than that in the breast muscle of Gushi chicken(P<0.05). The above results provide a theoretical basis of further exploring the biological function of chicken CEBPA gene in the lipid deposition process of chicken breast muscle.

Key words: Gushi chicken, CEBPA gene, bioinformatics analysis, breast muscle, expression analysis