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.
出处
《中国畜牧兽医》
CAS
北大核心
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)
