无量山乌骨鸡FSHR基因分离、表达及其蛋白功能分析

Isolation and Expression of FSHR Gene and Its Protein Function Analysis in Wuliangshan Silky Chicken

【目的】揭示无量山乌骨鸡FSHR基因的结构与功能。【方法】采用RT-PCR法克隆了无量山乌骨鸡FSHR基因的编码区全序列,并利用生物信息学方法对无量山乌骨鸡FSHR基因编码产物的理化特性、结构及功能进行了初步分析,最后采用实时荧光定量分析其组织表达情况。【结果】本研究获得的无量山乌骨鸡FSHR基因编码区全长为2 082 bp,编码693个氨基酸。生物信息学预测显示:无量山乌骨鸡FSHR无N-端信号肽。该蛋白含有3个保守结构域、7个跨膜区和6种功能活性位点。无量山乌骨鸡FSHR二级结构主要由无规则卷曲和α螺旋构成,分别占35.50%和31.17%。氨基酸序列比对显示:无量山乌骨鸡FSHR与其他禽类的同源性在87.4%以上。荧光定量结果显示:该基因在性腺组织中表达量最高,说明该基因与生产性能、繁殖性能等有关系。【结论】FSHR属于G蛋白偶联受体家族,可介导促卵泡作用的发生,可能在转运和结合、信号转导、修饰加工等过程中发挥重要作用。

[Purpose]Our aim was to reveal the structure and function of Wuliangshan silky chicken FSHR gene.[Method]The whole coding sequence(CDS)of the FSHR gene was cloned using RT-PCR method,its tissue expression was studied by real-time fluorescence quantitative analysis,and the physicochemical properties,structure and function on its coding product were analysed preliminarily based on the method of bioinformatics.[Results]The completed CDS of Wuliangshan silky chicken FSHR gene is 2 082 bp in length,encoding a peptide composed of 693 amino acid residues.Bioinformatics analysis showed that the Wuliangshan silky chicken FSHR has not a signal peptide.This protein contains three conservative domains,seven transmembrane domains and six kinds of functional active sites.The secondary structure of FSHR in Wuliangshan silky chicken are mainly composed of random curl andα-helix,occupied 35.50%and 31.17%respectively.The amino acid sequence alignment showed that homology is more than 87.4%between Wuliangshan silky chicken and other poultry.Fluorescent quantification results showed that the gene expression in gonadal tissue highest,indicating that the gene relative to the production performance,reproductive performance.[Conclusion]FSHR belongs to the G protein coupled receptor family,which is mediate follicularstimulating effects and may play an important role in transport and binding,signal transduction,modification and processing.