鸡Akirin同源基因的克隆与组织表达分析

Cloning and Tissue Expression Analysis of Chicken Akirin Homologous Gene

Akirin是最近被发现的,在果蝇和小鼠的免疫炎症反应和调控肌肉发生再生中起着重要作用的新基因.本实验采用电子克隆技术成功克隆获得了海兰褐鸡Akirin基因的全长编码序列.利用生物信息学方法对Akirin基因进行序列分析和预测,并利用半定量RT-PCR技术对雏鸡和成鸡的Akirin基因进行组织表达谱分析.结果表明,鸡Akirin同源基因可读框长576 bp,进化保守高,编码蛋白的氨基酸序列存在PFAM、RGS、TOP1Bc、UTG、HMG和low complexity sequence几种结构域,和蛋白激酶C磷酸化位点、酪氨酸激酶磷酸化位点和N端豆蔻酰化位点3个功能位点,同时在鸡Akirin的3′非翻译区预测到1个microRNA靶位点.Akirin在鸡的心、脾、脑等多种组织中广泛表达,推测鸡Akirin基因是一个可能具有多种功能作用的新基因.本研究将为深入研究鸡Akirin基因功能作用奠定基础.

The Akirin gene is a recently discovered to plays important roles in regulation of immune and inflammatory response,as well as in the myogenesis in Drosophila and mice.In this study,the full-length Akirin gene in Hi-Line Brown chicken was cloned using an in silico approach.The sequence homology and protein functions of chicken Akirin were predicted using bioinformatic analysis,and the tissue expression pattern was investigated using semi-quantitative RT-PCR in both the newly hatched and adult chicken.The full-length coding sequence of chicken Akirin gene is 576 bp and conserved evolutionally.The Akirin protein sequence is predicted to have several functional domains,including PFAM,RGS,TOP1Bc,UTG,HMG and some low complexity sequence.Three sites as the protein kinase C phosphorylation,tyrosine kinase phosphorylation,N-myristoylation can also be found in the amino acid sequence.One predicted microRNA targeting site was identified in the 3′ untranslated region of chicken Akirin mRNA.The Akirin expression was detected in a variety of organizations,such as heart,spleen,brain,etc.Our results suggested that chicken Akirin might be a novel gene with multiple functions.