绒山羊胎儿期毛囊发育相关microRNA的筛选与鉴定

发布时间：2018-11-05 19:33

【摘要】：内蒙古绒山羊具有优良的产绒性状,是宝贵的种质遗传资源,因此研究绒山羊的绒毛发育机理,探寻与绒毛生长相关的重要候选基因就显得尤为重要。本研究采集了绒山羊10个不同时期的胎儿皮肤,根据前期研究结果对绒山羊胎儿皮肤毛囊启动、生长发育3个关键时期45天、55天和65天的绒山羊胎儿的体侧皮肤(相同时期的3个样本当作生物学重复)的RNA样本,等量混合后进行高通量组测序,并对结果进行生物信息学分析及荧光定量PCR(qPCR)验证。试验结果显示：1、运用高通量测序方法获得绒山羊不同胎儿期皮肤毛囊microRNA和mRNA表达数据。oar-let-7和oar-miR-200家族的microRNA在妊娠55天和妊娠65天样本中,都相对于妊娠45天显著上调,暗示其可能是毛囊发育中关键的microRNA。2、应用生物信息学分析预测oar-let-7和oar-miR-200的靶基因为GLDC、GPM6A、LRP。3、试验采用荧光定量PCR技术对绒山羊胎儿期皮肤毛囊测序结果中关键的microRNA进行验证,结果显示定量结果表达趋势跟测序结果的基本一致。4、对预测的靶基因进行验证,45天、55天、65天、75天、85天、95天、105天、115天、125天、135天十个时期预测的三个靶基因GLDC、GPM6A、LRP均有表达,在一定程度上说明了这些靶基因对于调控绒山羊胎儿期皮肤毛囊发育有明显作用。
[Abstract]:Inner Mongolia cashmere goats are valuable germplasm genetic resources, so it is very important to study the mechanism of cashmere development and to search for important candidate genes related to the growth of cashmere in Inner Mongolia Cashmere Goat (Inner Mongolia Cashmere Goat). In this study, 10 fetal skin samples were collected from cashmere goats at different stages. According to the results of previous studies, the hair follicles of cashmere goats fetal skin were started, and the growth and development of fetal hair follicles were carried out for 45 days. RNA samples from the body side skin of the 55-day and 65-day cashmere goat fetuses (three samples from the same period as biological duplicates) were mixed in the same amount and sequenced in the high-throughput group. The results were analyzed by bioinformatics and verified by fluorescence quantitative PCR (qPCR). The results showed that: 1. The data of microRNA and mRNA expression in hair follicles of cashmere goats at different fetal stages were obtained by high-throughput sequencing. MicroRNA from oar-let-7 and oar-miR-200 families were collected on day 55 and day 65 of gestation. Both were significantly up-regulated relative to 45 days of pregnancy, implying that they may be key to hair follicle development by using bioinformatics analysis to predict the target bases of oar-let-7 and oar-miR-200 because of GLDC,GPM6A,LRP.3,. Fluorescence quantitative PCR technique was used to verify the key microRNA of hair follicle sequencing in fetal cashmere goat. The results showed that the trend of quantitative expression was basically the same as that of sequencing. 4. The predicted target gene was verified for 45 days. The three target genes GLDC,GPM6A,LRP were expressed in 55 days, 65 days, 75 days, 85 days, 95 days, 105 days, 115 days, 125 days and 135 days. To some extent, these target genes play an important role in regulating the development of hair follicles in cashmere goats.
【学位授予单位】：内蒙古农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S827

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