番鸭ERBB4基因组织表达、克隆及序列分析

Tissue Expression,Cloning and Bioinformatics Analysis of ERBB4 Gene in Muscovy Duck(Cairna moschata)

试验旨在检测受体酪氨酸蛋白激酶4(receptor tyrosine protein kinase 4,ERBB4)基因在番鸭不同生理状况下脑组织中的相对表达水平,分析其生物信息学功能,探究ERBB4基因是否影响鸭的攻击行为。运用实时荧光定量PCR方法对番鸭的纹状体、下丘脑中ERBB4基因的相对表达量进行检测,通过RT-PCR扩增并克隆ERBB4基因CDS核心区后测序,分析其同源性及遗传进化关系,并结合生物信息学分析工具预测ERBB4蛋白的理化特性、亚细胞定位、信号肽、二级结构、三级结构等。结果显示,ERBB4基因在番鸭两个脑组织中都有一定的表达,在攻击行为番鸭脑组织中的表达量高于正常组,在纹状体中的表达量略高于下丘脑。ERBB4基因核苷酸与氨基酸序列同源性比对显示,ERBB4基因在不同物种间具有一定的遗传多样性,与绿头鸭亲缘关系最近。ERBB4蛋白为亲水性稳定蛋白,氨基酸序列的N端不存在信号肽,存在5个富含呋喃样半胱氨酸(FU)结构域和1个酪氨酸激酶(TyrKc)蛋白结构域,亚细胞定位于细胞质;空间结构以右手α-螺旋和无规则卷曲为主。综上,ERBB4基因可能在番鸭神经系统中发挥重要作用,本研究结果为进一步深入探讨ERBB4基因在番鸭攻击行为中的作用机制提供了参考资料。

This study was aimed to detect the relative expression level of receptor tyrosine protein kinase 4(ERBB4)gene in brain under different physiological conditions of muscovy ducks,analyze its bioinformatics function,and explore whether ERBB4 gene affected the aggressive behavior of ducks.The relative expression of ERBB4 gene in striatum and hypothalamus of muscovy ducks was detected by Real-time quantitative PCR.The CDS core region of ERBB4 gene was cloned and sequenced by RT-PCR,analyze the homology and genetic evolution of ERBB4 gene,and predict the physicochemical properties,subcellular localization,signal peptide,secondary structure and tertiary structure by bioinformatics analysis tools.The results showed that ERBB4 gene was expressed in both brain tissues of muscovy ducks,the expression in brain of muscovy ducks with aggressive behavior was higher than that of normal group,and the expression in striatum was slightly higher than that of hypothalamus.The homology of nucleotide and amino acid sequences showed that ERBB4 gene had certain genetic diversity among different species and was closely related to Anas platyrhynchos.ERBB4 protein was hydrophilic and stable.There was no signal peptide at the N-terminal of the amino acid sequence,5 furan-like cysteine(FU)rich domains and 1 tyrosine kinase(TyrKc)protein domain.The subcellular localization was in cytoplasm,and the spatial structure was mainly right-handed alpha helix and random coil.In addition,ERBB4 gene might play an important role in the nervous system of muscovy ducks,this results provided a reference for further exploring the mechanism of ERBB4 gene in aggressive behavior of muscovy ducks.