【摘要】 羊驼是毛用型经济动物,其耳部和背部的毛发品质和生长速度存在差异.MicroRNA(miRNA)是新发现的一类在转录后水平调控基因表达的非编码RNA分子,为比较miRNA在羊驼耳部和背部皮肤的表达差异,从而探讨miRNA在羊驼皮肤和毛囊发育过程中的调控作用,本实验提取羊驼皮肤总RNA,制备了羊驼皮肤miRNA芯片,通过与Affymetrix多物种miRNA芯片跨物种杂交对耳部和背部皮肤的miRNA进行筛选,并通过实时荧光定量PCR进行了验证,同时利用在线生物信息软件预测miRNA靶基因.结果显示,羊驼耳部和背部皮肤中高表达差异2倍以上的miRNA有39个,实时荧光定量PCR检测let-7b和miR-24在2个部位皮肤中的差异表达量与miRNA基因芯片结果一致;预测到let-7b和miR-24的靶基因中包含有与毛囊生长发育和毛发品质相关的基因,提示这些miRNA可能参与羊驼皮肤和毛囊的生长发育、更新以及毛发品质的调控.更多还原

【Abstract】 Alpaca(Lama pacos) is a fiber-producing animal of commercial importance.The hair from the ear and back skin are different in both quality and growth.It was recently recognized that microRNAs(miRNAs) was able to regulate the gene expression at post-transcription level.Investigating the difference of miRNA expression between ear and back skin in the alpaca may help to understand the regulatory mechanism of the development of alpaca skin and hair follicles.We isolated the total RNA from alpaca ear and back skin subjected to the hybridization to Affymetrix multi-species miRNA microarrays.The differentially expressed miRNAs were identified and verified with real-time PCR.The prediction of possible target genes were conducted using an online bioinformatics software.The results showed that 39 of the highly expressed miRNAs,were expressed with more than two folds difference between ear and back skin of alpaca,including let-7b and miR-24 that later confirmed by real-time PCR.We postulated that some of the let-7b and miR-24 target genes might be related to hair-follicle growth and hair quality.Our data also indicated the importance of miRNAs in the regulation of the development and growth of skin and hair-follicle which might affect the determination of hair quality.更多还原