基于RNA-seq分析褪黑素对内蒙古绒山羊毛囊发育相关基因表达的影响

发布时间：2018-04-17 21:08

本文选题：绒山羊 + 褪黑激素　；参考：《内蒙古农业大学》2017年硕士论文

【摘要】：本研究基于课题组前期获得的内蒙古绒山羊皮肤组织的转录组数据,通过GO信息注释、KEGG信号通路分析,得到内蒙古绒山羊皮肤组织试验组与对照组2月、10月、12月的转录组数据。选取Wnt10b、β-catenin、SFRP1、FGF21、TCHHL1等5个差异基因作为候选基因,通过荧光定量PCR技术和Western-blot技术,分析这5个基因在mRNA水平和2个蛋白水平上的表达,为深入探索褪黑激素促进绒毛生长机理提供理论依据,主要研究结果如下:(1)应用生物信息学技术对内蒙古成年绒山羊试验组与对照组2月、10月、12月皮肤组织转录组数据进行差异基因的筛选。3个月份共筛选出1478个差异基因,其中上调的1014个,下调的464个。(2)荧光定量PCR结果显示:埋植褪黑激素,使Wnt10b基因在1月、3月表达量显著的高于对照组(P0.05),在2月的表达量显著低于对照组(P0.05);β-catenin基因在11月、1月表达量显著高于对照组(P0.05),在7月表达量显著低于对照组(P0.05);FG21基因在,11月、1月、2月表达量显著高于对照组(P0.05);SFRP1基因在6月、7月表达量显著高于对照组(P0.05),在其他月份均无显著差异(P0.05)TCHHL1基因在4月、6月、8月表达量显著低于对照组(P0.05),1月、2月的表达量显著高于对照组(P0.05)。(3)Western blot结果显示:Wnt1Ob蛋白的表达量在12月、2月试验组显著高于对照组(P0.05);而在9月差异不显著(P0.05)。β-catenin蛋白表达量在9月试验组显著低于对照组(P0.05),而在12月、2月试验组与对照组β-catenin蛋白表达量差异不显著(P0.05)。由以上研究结果可知,Wnt10b、β-catenin、SFRP1、FGF21、TCHHL1等5个差异基因在褪黑激素促进绒毛生长发育中起到重要作用。
[Abstract]:Based on the transcriptional data of Inner Mongolia Cashmere Goats (Cashmere Goats), we obtained the transcriptional data of Inner Mongolia Cashmere Goat skin tissue test group and control group in February, October and December through go information annotation and KEGG signal pathway analysis.Five differentially expressed genes, Wnt10b, 尾 -cateninine SFRP1FGF21, TCHHL1, were selected as candidate genes. The expression of these five genes at mRNA level and 2 protein levels were analyzed by fluorescence quantitative PCR and Western-blot techniques.It provides a theoretical basis for further exploring the mechanism of melatonin promoting the growth of villi.The main results are as follows: (1) using bioinformatics to screen differentially expressed genes between the experimental group of Inner Mongolia adult cashmere goat and the control group in February, October and December. A total of 1478 differentially expressed genes were screened in three months.Of these, 1014 were up-regulated and 464 were down-regulated. PCR results showed that melatonin was implanted.The expression of Wnt10b gene was significantly higher in January and March than that in control group, and was significantly lower in February than that in control group, while the expression of 尾 -catenin gene in November and January was significantly higher than that in control group, and in July it was significantly lower than that in control group.In November, January and February, the expression level of SFRP1 gene was significantly higher than that of control group in June, and the expression level in July was significantly higher than that in control group. There was no significant difference in other months. The expression of TCHHL1 gene in April, June and August was significantly lower than that in control group, 2 January.The results of Western blot showed that the expression of Wnt1Ob protein in the control group was significantly higher in December than that in the control group in 2 months, but not in September. The expression of 尾 -catenin protein in the control group was significantly lower than that in the control group in September, and the expression of 尾 -catenin protein in the control group was significantly lower than that in the control group in September, and the expression of 尾 -catenin protein in the control group was significantly lower than that in the control group in September.However, there was no significant difference in the expression of 尾 -catenin between the experimental group and the control group in 12 months and 2 months.According to the above results, five differentially expressed genes, Wnt10b, 尾 -cateninine SFRP1FGF21 and TCHHL1, play an important role in the growth and development of villi by melatonin.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S827

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