CAJ | 학술논문

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140 bp ,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,G nαq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 G nαq可能参与了小鼠被毛颜色的形成过程.
위료탐토조표령핵감산결합단백α아기q( guanine nucleotide binding protein alpha q , Gnαq)기인여동물모색형성적관계,채용반전록취합매련반응( reverse transcription‐polymerase chain reaction , RT‐PCR)、실시형광정량취합매련반응( quantitative real‐time polymerase chain reaction ,qRT‐PCR)기술화면역인적법( western blotting)검측 Gnαq mRNA화기단백질적상대표체량；이용면역조직화학법대Gnαq단백질재불동모색피부조직중진행정위분석.RT‐PCR결과현시：Gnαq적부분서렬피확증출,확증편단장도위140 bp ,경측서소득위목적편단.장소득기인서렬재NCBI상여기타물충적서렬진행동원성비대후발현,소서적 Gnαq서렬여기타물충적Gnαq서렬유교고적동원성,우기여대서、회창서적동원성최고,분별위98％화92％.qRT‐PCR화면역인적결과현시：재흑색소서피부조직중 Gnαq mRNA표체량화Gnαq단백질표체량균겁현저고우백색소서피부조직( P＜0.01).면역조직화학법검측표명,Gnαq단백질재백색화흑색소서모낭중적세포주위표체,차재모낭적외근초화모구부유표체.총지,G nαq재불동모색피부조직중적표체량유일정적차이,차분포재모낭적외근초화모구부,설명 G nαq가능삼여료소서피모안색적형성과정.
Summary Rich coat color is not only an important biological characteristics , but also a significant economic value for wool producing animals . Guanine nucleotide binding protein alpha q (Gnαq) is a downstream subunit of heterologous trimeric G protein . It plays a significant role in the performance of endothelin B receptor , which is involved in regulating coat hair of animals . <br> In order to understand how Gnαq 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 Gnαq gene in different mice with two kinds of colors (white and black) . The localizations of Gnαq protein were analyzed by immunohistochemistry in different color skin tissues of mice . <br> A partial sequence of Gnαq gene with the length of 140 bp was successfully amplified by PCR . Blasting in NCBI showed that the Gnαq 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 Gnαq mRNA and Gnαq protein in black skin of mice was significantly higher than those in the w hite skin of mice ( P< 0 .01 ) . According to the results of immunohistochemistry , the Gnαq 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 . <br> In conclusion , the expression of Gnαq 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 Gnαq gene may play a critical role in the process of coat color formation in mice .