猪NR4A1基因的克

发布时间：2018-01-07 13:23

本文关键词：猪NR4A1基因的克隆、序列分析及表达模式研究　出处：《西北农林科技大学学报(自然科学版)》2016年01期 　论文类型：期刊论文

【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。
[Abstract]:[objective] to clone nuclear receptor subfamily group member 1 from porcine nuclear receptor 4A1. The NR4A1)DNA sequence was full-length, and the CDS sequence and its expression pattern were analyzed. [methods] the ovaries of adult sows were used as materials, and the technique of electronic cloning was used. The porcine NR4A1DNA sequence was isolated and sequenced, and the porcine NR4A1 gene sequence was analyzed by bioinformatics. Human chorionic gonadotropin was used as human chorionic gonadotropin. HCG) and Pregnant mare serum gonadotropin (PMSG) were used to treat ovarian granulosa cells. The expression pattern of porcine NR4A1 gene was analyzed by RT-PCR method. [results] the DNA sequence of porcine NR4A1 gene was 4870 BP. The full-length CDS sequence was 1 734 BP, encoding 572 amino acids. The amino acid sequence encoded by porcine NR4A1 gene was similar to the corresponding amino acid sequence of human being, and the highest similarity was found between the amino acid sequence of pig NR4A1 gene and the corresponding amino acid sequence of human. The predicted results of Expasy software showed that the amino acid sequence of NR4A1 protein was very conserved, which contained two zinc finger features NR C4-type, one of which was a hormone receptor superfamily. The other is the Zf-C4 superfamily; Porcine ovarian granulosa cells treated with gonadal hormone could induce the transient expression of porcine NR4A1 gene during follicular development and ovulation. [conclusion] the CDS region of porcine NR4A1 gene is more conserved among species. It is inferred that porcine NR4A1 gene regulates follicle development and ovulation.
【作者单位】：安徽省农业科学院畜牧兽医研究所;华中农业大学动物科学院;
【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)
【分类号】：S828
【正文快照】： NR4A1基因作为一个孤核受体的转录因子,编码类固醇-甲状腺激素超家族。NR4A1蛋白是一类配体依赖转录调节蛋白[1],它在转录过程中起作用的是N-末端,参与调节核定位和转录激活过程的是C-末端[2-4]。在细胞中进行的类固醇合成活性研究结果显示,NR4A1基因表达与P450胆固醇侧链裂解

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