Syntaxin 17在不同毛色小鼠皮肤中差异表达与在毛囊中的定位

发布时间：2018-04-24 01:27

本文选题：突触融合蛋白17 + 不同毛色　；参考：《山西农业大学》2015年硕士论文

【摘要】：目的：动物毛纤维是人类生产生活不可或缺的重要资源,主要毛用性状有细度、长度和色彩。而毛色作为主要毛用性状之一,是由许多基因共同决定的。老鼠作为重要的实验动物,对其毛色的研究有助于对于其他动物毛色的研究。方法：为了研究Stx17基因与小鼠毛色形成中的相关性,本试验通过在小鼠皮肤组织和小鼠的黑色素细胞上扩增的Stx17基因的序列,并用荧光定量实时PCR、免疫化学和Western blot对其在不同毛色皮肤组织中进行定量和定性的分析,以说明Stx17基因在不同毛色的小鼠的皮肤组织上的表达差异,从而为进一步研究Stx17基因在不同毛色形成过程中的作用机制打好基础。结果：(1)通过PCR技术成功扩增出Stx17基因全cds区序列,经测序所得为目的片段,在NCBI上与其他物种进行同源序列的比对,得知与其他物种的基因有较高的同源性。(2)通过Real-Time PCR研究stx17基因在不同毛色小鼠皮肤组织中的相对表达量,结果显示白鼠和灰腹表达水平分别是黑背的0.115倍和1.682倍,差异极显著(P0.01),可见野生型小鼠的灰色腹部中的基因表达量显著高于白色小鼠个体和黑色背部,这说明该基因与灰色的形成有一定的相关性。(3)应用免疫印迹技术(Western Blotting)对STX17蛋白在不同毛色皮肤组织中的蛋白表达进行研究,结果显示在白色小鼠、灰色和黑色小鼠皮肤组织中均有蛋白的表达,并且在灰色小鼠皮肤组织中的表达量是最高的。(4)通过免疫化学法观察Stx17基因在小鼠皮肤组织中的定位,结果显示Stx17基因在小鼠皮肤的真皮部位呈阳性表达。并对小鼠的黑色素细胞进行免疫化学染色后也证实STX17在黑素细胞的细胞膜上有表达。结论：以上所述表明,Stx17基因在不同毛色的皮肤组织中都有表达,而且表达量有一定的差异性,因此,推测该基因可能与不同毛色的形成具有相关性。
[Abstract]:Objective: animal hair fiber is an indispensable resource for human production and life. The main wool characters are fineness, length and color. As one of the main wool traits, coat color is determined by many genes. As an important laboratory animal, the study of the hair color of mice is helpful to the study of other animal's coat color. Methods: in order to study the correlation between Stx17 gene and mouse coat color formation, the Stx17 gene sequence was amplified from mouse skin and melanocytes. Fluorescence quantitative real-time PCR, immunocytochemistry and Western blot were used to quantitatively and qualitatively analyze the expression of Stx17 gene in the skin tissues of mice with different coat colors, so as to explain the difference in the expression of Stx17 gene in the skin tissues of mice with different coat colors. The results provide a basis for further study on the mechanism of Stx17 gene in the process of different coat color formation. Results the whole cds region of Stx17 gene was successfully amplified by PCR. The sequence was sequenced and compared with other species on NCBI. The results showed that the relative expression of stx17 gene in the skin of different hairy mice was 0.115 times and 1.682 times higher than that of black back, respectively, and the expression level of stx17 gene was 0.115 times and 1.682 times higher than that of black back, respectively, by using Real-Time PCR, the results showed that the expression level of stx17 gene was 0.115 times and 1.682 times higher than that of black back, respectively. The difference was very significant (P 0.01). It was found that the gene expression in gray abdomen of wild type mice was significantly higher than that of white mice and black back. The results showed that the expression of STX17 protein in different skin tissues was studied by Western blotting technique. The results showed that the expression of STX17 protein was found in white mice. The expression of protein was found in both gray and black mouse skin tissues, and the highest expression level was found in gray mouse skin tissues.) Immunochemical method was used to observe the localization of Stx17 gene in mouse skin tissues. The results showed that Stx17 gene was positively expressed in the dermis of mice. Immunocytochemical staining of mouse melanocytes also confirmed that STX17 was expressed on melanocyte membrane. Conclusion: these results suggest that the Stx17 gene is expressed in skin tissues with different coat colors, and the expression levels are different. Therefore, it is speculated that this gene may be related to the formation of different coat colors.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.2

【参考文献】