辽宁新品系绒山羊GMPR2基因的生物学功能研究  

Study on Biological Function of GMPR2Gene in Liaoning New-breeding Cashmere Goat

作　　者：金梅[1] 张丽丽[1] 曲杨乐[1] 朴君[1] 李秀娜[1] 王宜萌 朴敬爱[1] 

机构地区：[1]辽宁师范大学生命科学院,辽宁省生物技术与分子药物研发重点实验室,大连116029

出　　处：《畜牧兽医学报》2013年第11期1706-1716,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31172188);大连市科技计划项目(2013B12NC090);辽宁省百千万人才基金联合资助

摘　　要：本研究在辽宁新品系绒山羊中发现了GMPR2基因,旨在研究其在皮肤中的功能作用与绒毛生长的关系。首先对此基因进行了生物信息学分析,再利用地高辛标记的原位杂交技术对其进行定位分析,然后利用荧光定量技术对其进行相对定量分析,RT-PCR技术检测GMPR2基因是否在其他器官中表达,最后验证不同浓度褪黑激素处理不同时间是否影响其表达。结果表明,得到的基因全长确为GMPR2基因,与牛GMPR2同源性最高;在初次级毛囊中都表达,表达部位为内根鞘;GMPR2基因在退行期初级毛囊中的表达量最高(P<0.01),在兴盛期与休止期表达量相差不明显;另外,GMPR2基因只在皮肤中表达,在心、肝等内脏器官不表达;褪黑激素处理后,GMPR2的表达量明显降低。所以,推断GMPR2可能间接调节初、次级毛囊的生长萎缩。This study aimed to investigate the function of GMPR2 gene which was found in the skin of Liaoning New-breeding Cashmere goat and its relationship with the cashmere fiber growth. Bioinformatics analysis was firstly used, and then positioning analysis was got by using DIG-labeling in situ hybridization technique, and semi-quantitative real-time PCR was used to de- tect the levels of GMPR2 gene expression in different periods, and then, the RT-PCR was used to detect whether GMPR2 gene was expressed in other organs. Finally, gene expression was affected by different concentration melatonin treatment at different time. The results showed that the gene was really GMPR2 gene, which showed high homology to the Bos taurus GMPR2 gene. The in situ hybridization result showed that GMPR2 had a strong signal in inner root sheath of primary and secondary follicles. Besides, the expression of GMPR2 genes in primary follicles dur ing catagen was the highest （P^0.01）, but the expression was not obviously different between anagen and telogen. In addition, RT-PCR analysis showed that GMPR2 gene was only expressed in goat skin, not in liver and heart. The expression quantity of GMPR2 was decreased after me latonin treatment. It was infered that GMPR2 may indirectly regulate the primary and secondry follicles stunted growth.

关 键 词：GMPR2基因 生物信息学分析 原位杂交 荧光定量 RT—PCR 褪黑激素 

分 类 号：S827[农业科学—畜牧学] S813.1[农业科学—畜牧兽医]