黄麻羽肉鸡β-防御素基因AvBD1和AvBD2克隆、生物信息学分析及组织表达

doi:10.16431/j.cnki.1671-7236.2023.11.002

摘要/Abstract

摘要： 【目的】防御素是生物体内产生的具有广谱抗微生物作用的活性肽。深入认识鸡β-防御素AvBD1和AvBD2的基本信息，有助于进一步研究和应用其生物学功能。【方法】采用PCR方法扩增黄麻羽肉鸡AvBD1和AvBD2基因并克隆测序，将测序获得的CDS序列推导成氨基酸序列后进行相似性比对，利用在线软件分析所预测蛋白的生物信息学特性，并探究二者在黄麻羽肉鸡不同组织中的表达情况。【结果】黄麻羽肉鸡AvBD1和AvBD2基因CDS区大小分别为198和195 bp，可编码65和64个氨基酸，二者CDS区序列相似度为56%，氨基酸序列相似度为36%，可见二者并非同源基因。系统进化树表明，黄麻羽肉鸡与其他几个品种鸡的亲缘关系较近，其中AvBD1基因与小鼠亲缘关系最远；AvBD2基因与大山雀亲缘关系最远。生物信息学分析结果显示，AvBD1和AvBD2均为疏水性蛋白，且AvBD1疏水性和稳定性均强于AvBD2，均存在信号肽，但所处氨基酸位点不同，无跨膜结构域，二级结构主要元件分别为α-螺旋和无规则卷曲，但参与形成这2种构象的氨基酸比例差异较大。实时荧光定量PCR分析发现，AvBD1和AvBD2基因在黄麻羽肉鸡心脏、肝脏、肺脏、肾脏和十二指肠组织中整体表达趋势相似，均以肺脏中表达量最高，十二指肠中表达量最低。【结论】试验成功克隆了黄麻羽肉鸡AvBD1和AvBD2基因CDS区全长序列，分析了AvBD1和AvBD2蛋白的结构和功能，二者在黄麻羽肉鸡内脏组织中的mRNA表达趋势一致，为进一步阐明AvBD1和AvBD2的生物学功能及其在生产中的应用提供参考依据。

关键词: 黄麻羽肉鸡; AvBD1基因; AvBD2基因; 克隆; 序列分析; 组织表达

Abstract: 【Objective】 Defensins are novel active peptides produced in vivo with broad spectrum antimicrobial activity.In-depth understanding of the basic information of β-defensins AvBD1 and AvBD2 in chickens will be helpful for further study and full application of their biological functions.【Method】 The sequences of AvBD1 and AvBD2 genes in Yellow-feather chickens were amplified using PCR, then cloned and sequenced, subsequently the CDS sequences obtained by sequencing were deduced as amino acid sequences for similarity alignment.The bioinformatics characteristics of the predicted proteins were analyzed by online software, and the expression of the corresponding encoding genes in different tissues of Yellow-feather chickens were compared.【Result】 The CDS length of AvBD1 and AvBD2 genes in Yellow-feather chickens were 198 and 195 bp, which encoded 65 and 64 amino acids, respectively.The similarity of CDS between AvBD1 and AvBD2 genes was 56%, and the similarity of amino acid sequences was 36%, indicating that the two were not homologous genes.The phylogenetic tree showed that Yellow-feather chickens had the closer genetic relationship with several other breeds of chickens, and AvBD1 and AvBD2 genes had the furthest genetic relationship with Mus musculus and Parus major, respectively.Bioinformatics analysis displayed that both AvBD1 and AvBD2 were hydrophobic proteins, and AvBD1 had stronger hydrophobicity and stability than AvBD2.They all contained signal peptides, however, they were located at different amino acid sites and there was no transmembrane domain.The secondary structure elements were mainly alpha helix and random coil, respectively, but the proportion of amino acids involved in the formation of the two conformations was significantly different.Real-time quantitative PCR results found that the overall expression trend of AvBD1 and AvBD2 genes were similar in heart, liver, lung, kidney and duodenum of Yellow-feather chickens, with the highest expression in lung and the lowest expression in duodenum.【Conclusion】 In this study, the CDS sequences of AvBD1 and AvBD2 genes in Yellow-feather chickens were cloned successfully, and the structure and function of the corresponding proteins were analyzed.Both of them were consistent at the mRNA level in visceral tissues of Yellow-feather chickens.The results laid a reference for further elucidating the biological function and application of AvBD1 and AvBD2 in production.

Key words: Yellow-feather chickens; AvBD1 gene; AvBD2 gene; cloning; sequence analysis; tissue expression

中图分类号:

Q78
S831

康丹菊, 彭秋云, 周嘉欣, 陈进军, 林红英, 周光现. 黄麻羽肉鸡β-防御素基因AvBD1和AvBD2克隆、生物信息学分析及组织表达[J]. 中国畜牧兽医, 2023, 50(11): 4325-4337.

KANG Danju, PENG Qiuyun, ZHOU Jiaxin, CHEN Jinjun, LIN Hongying, ZHOU Guangxian. Cloning, Bioinformatics Analysis and Tissue Expression of AvBD1 and AvBD2 Genes in Yellow-feather Chickens[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4325-4337.

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