山羊CXCR1基因的克隆及组织表达谱研究

Cloning and tissue expression profile of CXCR1 gene in goat

为了探究山羊C-X-C趋化因子受体1(C-X-C motif chemokine receptor 1,CXCR1)基因的特点,试验以金堂黑山羊脾脏cDNA为模板克隆了CXCR1基因,借助生物信息学软件及在线软件分析序列,并采用实时荧光定量PCR方法检测CXCR1基因在金堂黑山羊脾脏、肺脏、肌肉、心脏和肠的5个组织中的相对表达量。结果表明:克隆得到的CXCR1基因序列全长为1 140 bp,完整CDS编码区长1 107 bp,共编码368个氨基酸;所得氨基酸序列与牛进行比对,两者同源性为96.8%;编码蛋白为带正电荷、具有疏水性的稳定蛋白;有7个跨膜结构和32个磷酸化位点,无信号肽;二级结构中α-螺旋比例最高,为54.35%;获得的三级结构模型与人的CXCR1蛋白(PDB:2lnl.1.A)同源性最高,为81.49%;氨基酸比对结果与系统进化树结果相近,金堂黑山羊与牛的亲缘关系最近;CXCR1基因主要表达于肌肉和心脏。说明CXCR1基因可能在山羊肌肉发育相关的信号通路中发挥作用。

In order to explore the characteristics of the C-X-C chemokine receptor 1(CXCR1)gene in Jintang black goat,the CXCR1 gene was cloned using Jintang black goat spleen cDNA as a template in this experiment.The obtained sequence was then analyzed by bioinformatics software and online software.The relative expression of CXCR1 gene in the spleen,lung,muscle,heart and intestine of Jintang black goat was detected by real time quantitative PCR.The results showed that the total length of the CXCR1 gene sequence was 1 140 bp,the complete CDS was 1 107 bp and was encoded by 368 amino acids.The obtained amino acid sequence was compared with cattle and the homology was 96.8%.The coding protein is a stable protein with positive charge and hydrophobicity.It has 7 transmembrane structures and 32 phosphorylation sites,but there is no signal peptide.The highest ratio of secondary structure wasα-helix(54.35%).The obtained CXCR1 tertiary structure model has the highest homology with human proteins(PDB:2 lnl.1.A),which is 81.49%.The results of amino acid alignment were similar to those of phylogenetic trees,indicating that Jintang black goat had the closest relationship with cattle.The CXCR1 gene was mainly expressed in muscles and hearts.The results indicated that the CXCRI gene may play a role in signal pathways related to muscle development in goats.