摘要
为了探讨鸟嘌呤核苷酸结合蛋白α亚基q(guanine nucleotide binding protein alpha q,Gnαq)基因与动物毛色形成的关系,采用反转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)、实时荧光定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)技术和免疫印迹法(western blotting)检测Gnαq mRNA和其蛋白质的相对表达量;利用免疫组织化学法对Gnαq蛋白质在不同毛色皮肤组织中进行定位分析。RT-PCR结果显示:Gnαq的部分序列被扩增出,扩增片段长度为140bp,经测序所得为目的片段。将所得基因序列在NCBI上与其他物种的序列进行同源性比对后发现,小鼠的Gnαq序列与其他物种的Gnαq序列有较高的同源性,尤其与大鼠、灰仓鼠的同源性最高,分别为98%和92%。qRT-PCR和免疫印迹结果显示:在黑色小鼠皮肤组织中Gnαq mRNA表达量和Gnαq蛋白质表达量均极显著高于白色小鼠皮肤组织(P<0.01)。免疫组织化学法检测表明,Gnαq蛋白质在白色和黑色小鼠毛囊中的细胞周围表达,且在毛囊的外根鞘和毛球部有表达。总之,Gnαq在不同毛色皮肤组织中的表达量有一定的差异,且分布在毛囊的外根鞘和毛球部,说明Gnαq可能参与了小鼠被毛颜色的形成过程。
Summary Rich coat color is not only an important biological characteristics, hut also a significant economic value for wool producing animals. Guanine nucleotide binding protein alpha q （Gnaq） is a downstream subunit of heterologous trimerie G protein. It plays a significant role in the performance of endothelin B receptor, which is involved in regulating coat hair of animals, In order to understand how Gnaq gene acts in the process of manipulating mammalian hair color, reverse transcription-polymerase chain reaction （RT-PCR）, quantitative real-time polymerase chain reaction （qRT-PCR） and western blotting were used to detect the difference, the relative expression of Gnaq gene in different mice with two kinds of colors （white and black）. The localizations of Gnaq protein were analyzed by immunohistochemistry in different color skin tissues of mice. A partial sequence of Gnc^q gene with the length of 140 bp was successfully amplified by PCR. Blasting in NCBI showed that the Gnc^q sequence of mice had a high homology with other species, especially with rat and gray dwarf hamster, with the similarity of 98% and 92%, respectively, qRT PCR and western blotting results showed that the relative expression of Gnaq mRNA and Gnaq protein in black skin of mice was significantly higher than those in the white skin of mice （P〈0.01）. According to the results of immunohistochemistry, the Gnaq protein was expressed around the cells in hair follicles of mice, as well as in the outer root sheath and hair bulb of hair follicle. In conclusion, the expression of Gnaq is significantly different between the white and black skin tissues of mice, and it is distributed in the outer root sheath and hair bulb of hair follicle, suggesting that the Gnaq gene may play a critical role in the process of coat color formation in mice.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2015年第4期414-420,共7页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
国家自然科学基金(30972223
31272628)
山西省研究生教育创新项目(2015BY30)
