绒山羊70d胚胎皮肤小分子RNA文库构建及miRNA鉴定  

Construction of a Small RNA Library and Identify of miRNAs from 70 Days Cashmere Goat Fetal Skin

作　　者：樊凯军[1] 尹俊[1] 

机构地区：[1]内蒙古农业大学生命科学学院,内蒙古呼和浩特010018

出　　处：《生物技术》2014年第1期32-36,共5页Biotechnology

基　　金：国家自然科学基金项目("BMP信号在绒山羊绒毛再生周期波中调节干细胞激活的研究";30960244)资助

摘　　要：目的：miRNA是一类21～23个核苷酸的单链小分子RNA，最近发现miRNA对哺乳动物毛囊的发育和维持具有重要作用。为了研究miRNA在绒山羊次级毛囊发育中的作用，可以为人工调节绒毛生长和提高羊绒产量提供新的线索。方法：该文构建了绒山羊70d胎儿胚胎体测部皮肤小RNA文库，库容量为2．5×10^7。随机挑选192个克隆提取质粒进行序列分析，序列经GenBank Blastn和mirBase搜索比对。结果：结果显示有32个序列和绒山羊及其它动物的miRNA高度同源，可以确定是山羊的miRNA。这些miRNA代表19个不同的miRNA序列，其中9个已注册，其余10个代表新的绒山羊miRNA。在克隆的miRNA中，miR-199a＊和miR-424的拷贝数最高，分别达到6个和3个，表明它他们在绒山羊次级毛囊发育期间的皮肤中非常活跃，可能有重要的生物学功能。结论：从绒山羊70d胚胎皮肤中鉴定了19个miRNA，这些miRNA可能参与绒山羊次级毛囊发育的调控。Objective：Recently, the miRNAs were found that plays an important role on of the hair follicle development and the mainte- nance. Study the role of the miRNAs during secondary hair follicles development of the cashmere goat ,in order to provide new clues for reg- ulating the cashmere growth and improving the cashmere production. Method：A small RNA library is constructed from the 70 - days cash- mere goat fetal skin,storage capacity is 2. 5 × 10^7. then the plasmids are extracted from 192 randomly selected clones for sequencing. These sequences are screening and comparing in GenBank and mirBase by Blastn. Result：The result indicated that 32 sequences are found which are highly homologous to other animal miRNAs. These miRNAs classify into 19 different miRNAs,miR - 199a ＊ and miR -424 have the maximum number of copies, respectively 6 and 3 and indicate they are very active during the hair follicle development of cashmere goat. Conclusion：This study is a foundation for research the role of these miRNAs during the secondary hair follicles development of cash- mere goat.

关 键 词：绒山羊 小分子RNA文库 MIRNA 次级毛囊 发育 

分 类 号：S818.9[农业科学—畜牧学]