猪NR4A1基因的克隆、序列分析及表达模式研究  

Cloning,sequence analysis and expression pattern of porcine NR4A1 gene

作　　者：刘林清[1] 李凤娥[2] 熊远著[2] 邓昌彦[2] 

机构地区：[1]安徽省农业科学院畜牧兽医研究所,安徽合肥230031 [2]华中农业大学动物科学院,湖北武汉430070

出　　处：《西北农林科技大学学报（自然科学版）》2016年第1期14-18,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：安徽省农业科学院科技创新团队项目(13C0405);安徽省农业科学院院长青年基金项目(14B0427)

摘　　要：【目的】克隆猪核受体4A1(Nuclear receptor subfamily 4,group A,member 1,NR4A1)DNA序列全长,并对CDS序列及其表达模式进行分析。【方法】以未成年长大母猪的卵巢为材料,采用电子克隆技术,对猪NR4A1DNA序列进行分离和测序,并对猪NR4A1基因序列进行生物信息学分析;利用性腺激素(人绒毛膜促性腺激素(Human chorionic gonadotropin,HCG)和孕马血清促性腺激素(Pregnant mare serum gonadotropin,PMSG))处理卵巢颗粒细胞,采用RT-PCR方法分析猪NR4A1基因的表达模式。【结果】获得猪NR4A1基因4 870bp的DNA序列,其中CDS全长序列为1 734bp,共编码572个氨基酸;猪NR4A1基因编码的氨基酸序列与人的对应氨基酸序列的相似性最高(97%),其次为小鼠(94%);Expasy软件预测结果表明,NR4A1蛋白的氨基酸序列非常保守,其中包含2个锌指特征(NR C4-type),一个是激素受体超家族,另一个是Zf-C4超家族;猪卵巢颗粒细胞在性腺激素处理后,可诱导猪NR4A1基因在卵泡发育和排卵时期瞬时表达。【结论】猪NR4A1基因的CDS区在物种间保守性较强,推测猪NR4A1基因调节卵泡的发育和排卵过程。[Objective] The genornic sequence of porcine NR4A1 gene was cloned and the temporal and spatial expression patterns were studied. [Method] Using ovary of minor sow as material, the structure of the porcine NR4A1 gene was analyzed by bioinformatics methods. The expression patterns of NR4A1 gene were analyzed by RT-PCR after ovarian granulosa cells were treated with pregnant mare serum gonadotro pin （PMSG） and human chorionic gonadotropin （HCG）. [Result] The genomic sequence with 4 870 bp of NR4A1 gene was obtained,which contained 1 734 bp CDS and encoded 572 amino acids. The amino acid se quences exhibited the greatest identity with human （97%）,followed by house mouse （94%）. Expasy soft ware estimation showed that NR4A1 protein was conservative including 2 NR （24 type,one was hormone receptor superfamily and the other was Zf-C4 superfamily. The porcine NR4A1 gene had a rapid expression pat^ern after granulose cells were treated with PMSG/HCG. [Conclusion] NR4A1 gene was highly conser vative among species and it involved in the regulation of follicular development,ovulation and e luteinizing process.

关 键 词：猪 NR4A1 序列分析 表达模式 

分 类 号：Q785[生物学—分子生物学]