基于UniGene寻找山羊新的miRNA及其试验验证  

Identification and Experimental Verification of Novel miRNA on UniGene in Goat

作　　者：苏蕊[1] 吕晓曼[1] 赵德超[1] 韩志玲[1] 付绍印[1] 孟丽云[1] 张文广[1] 李金泉[1] 

机构地区：[1]内蒙古农业大学动物科学学院/动物遗传育种与繁殖自治区重点实验室,内蒙古呼和浩特010018

出　　处：《中国畜牧兽医》2013年第11期7-11,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家科技支撑项目(2011BAD28B05);国家自然科学基金(30960246);内蒙古自治区自然科学基金(2011BS0407)

摘　　要：根据miRNA进化的保守性,以已知动物的miRNA为搜索序列与山羊的UniGene进行BLAST比对,预测山羊新的miRNA,并通过RT-PCR和克隆测序进行验证。对新发现的山羊miRNA进行实时荧光定量,检测其在12个月皮肤及肌肉中的表达量;用基于UniGene的方法获得了5条山羊miRNA候选分子,通过RT-PCR和克隆测序验证,发现这5条miRNA分子在山羊的皮肤和肌肉中均有表达,其中由chi-miR-374b、chi-miR-421和chi-miR-421*构成了山羊miRNA基因簇,chi-miR-2284n是牛和山羊特异的。实时荧光定量结果显示,chi-miR-1839、chi-miR-374b和chi-miR-2284n在2月份皮肤中的表达量明显高于其他月份,chi-miR-421和chi-miR-421*在10月份皮肤中的表达量最高;发现了5条山羊新的miRNA(chi-miR-2284n、chi-miR-421*、chi-miR-421、chi-miR-1839和chi-miR-374b,登录号:JQ002550-JQ002554),补充了山羊miRNA数据库的不足。Using goat UniGene to identify miRNA and RT-PCR was used to validate the predicted miRNA, and then Real- time PCR was performed to detect the miRNA expression during one year in goat skin and muscle. Results revealed that 5 new miRNA were identified and all of these miRNA were expressed in goat skin and muscle, in which a miRNA cluster was discow ered which was composed of chi-miR-421 ^＊ ,chi-miR-421 and chi-miR-374b, miRNA expression results indicated that chi-miR- 1839,chi-miR-374b and chi-miR-2284n in skin gave a highest expression level on February, chi-miR-421 and chi-miR-421 ^＊ ex- pression increased remarkably on October. The identification of 5 new miRNA in goat （chi-miR-2284n, chi-miR-421 ^＊ , chi- miR-421, chi-miR-1839 and chi-miR-374b,accession number：JQ002550-JQ002554） was a useful complement for goat miRNA database.

关 键 词：UNIGENE MIRNA 山羊 皮肤 肌肉 

分 类 号：Q78[生物学—分子生物学]