绵羊FGF10基因克隆、序列分析及其在毛囊发育中的表达分析

本试验旨在获得中国美利奴羊成纤维细胞生长因子10（fibroblast growth factor 10，FGF10）基因的编码区（CDS）全长序列并进行生物信息学分析，随后对FGF10基因在中国美利奴羊毛囊发育过程中的表达特征进行分析，明确其在中国美利奴羊毛囊发育过程中的表达模式，为进一步研究FGF10 mRNA表达水平与中国美利奴羊毛囊生长发育的表达调控机制奠定理论基础。采用PCR扩增获得中国美利奴羊FGF10基因CDS，并克隆到zero PCR@^TM-Blunt进行测序验证；利用实时荧光定量PCR技术检测FGF10在中国美利奴羊毛囊发育过程中的表达差异。结果表明，绵羊FGF10基因CDS长度为696 bp（序列上传GenBank，获得登录号：MT872422），编码231个氨基酸，与牛和山羊的氨基酸序列同源性达100%，存在1个信号肽和1个跨膜结构域，其为分泌通路信号蛋白；实时荧光定量PCR分析表明，FGF10基因在中国美利奴羊毛囊发育过程中均表达，在毛囊发育第85天表达最高，显著高于其他毛囊发育时期（P<0.05）。本研究获得中国美利奴羊FGF10基因完整的编码区序列和毛囊发育过程中的表达特征，生物信息学分析发现，FGF10基因编码区序列具有物种间的保守性，同时FGF10在绵羊毛囊不同发育阶段的皮肤组织中表达，由此表明，FGF10基因可能在绵羊毛囊的生长发育过程中发挥重要的生物学作用。

Cloning,Sequencing and Quantitative Expression of FGF 10 Gene in Sheep During Hair Follicle Development Period

The aim of this study was to obtain Chinese Merino sheep full-length sequence of the coding region(CDS)of fibroblast growth factor 10(FGF10)gene,to analyze its sequence characteristics by bioinformatics method,and to study the expression characteristics of FGF10 gene.It could lay a theoretical foundation for further study of the expression and regulation mechanism of FGF10 mRNA in Chinese Merino hair follicle.The full CDS of FGF10 gene was obtained by PCR amplification and cloned into zero PCR@TM-Blunt for sequencing verification.Real-time quantitative PCR(qRT-PCR)was used to detect the expression pattern of FGF10 gene.The results showed that the sheep FGF10 CDS was 696 bp in length(Upload the sequence to GenBank and get the accession No.:MT872422),which coded 231 amino acids,the homology of amino acid sequence with cattle and goat was 100%,there was a signal peptide and a transmembrane domain,which was the signal protein of secretory pathway.qRT-PCR analysis showed that FGF10 gene was expressed during the development of Chinese Merino follicles,and the highest expression was found on the 85th day of hair follicle development,which was significantly higher than those of other hair follicle development stages.The complete CDS and expression characteristics of FGF10 gene were obtained.Bioinformatics analysis showed that the CDS of FGF10 gene was conserved among species,and the gene was expressed in the skin tissue of different development periods of hair follicle,which indicated that FGF10 gene might play an important role in the growth and development of sheep hair follicles physical function.