本文關(guān)鍵詞：豬NR4A1基因的克隆、序列分析及表達(dá)模式研究　出處：《西北農(nóng)林科技大學(xué)學(xué)報(bào)(自然科學(xué)版)》2016年01期 　論文類(lèi)型：期刊論文

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【摘要】：【目的】克隆豬核受體4A1(Nuclear receptor subfamily 4,group A,member 1,NR4A1)DNA序列全長(zhǎng),并對(duì)CDS序列及其表達(dá)模式進(jìn)行分析�！痉椒ā恳晕闯赡觊L(zhǎng)大母豬的卵巢為材料,采用電子克隆技術(shù),對(duì)豬NR4A1DNA序列進(jìn)行分離和測(cè)序,并對(duì)豬NR4A1基因序列進(jìn)行生物信息學(xué)分析;利用性腺激素(人絨毛膜促性腺激素(Human chorionic gonadotropin,HCG)和孕馬血清促性腺激素(Pregnant mare serum gonadotropin,PMSG))處理卵巢顆粒細(xì)胞,采用RT-PCR方法分析豬NR4A1基因的表達(dá)模式。【結(jié)果】獲得豬NR4A1基因4 870bp的DNA序列,其中CDS全長(zhǎng)序列為1 734bp,共編碼572個(gè)氨基酸;豬NR4A1基因編碼的氨基酸序列與人的對(duì)應(yīng)氨基酸序列的相似性最高(97%),其次為小鼠(94%);Expasy軟件預(yù)測(cè)結(jié)果表明,NR4A1蛋白的氨基酸序列非常保守,其中包含2個(gè)鋅指特征(NR C4-type),一個(gè)是激素受體超家族,另一個(gè)是Zf-C4超家族;豬卵巢顆粒細(xì)胞在性腺激素處理后,可誘導(dǎo)豬NR4A1基因在卵泡發(fā)育和排卵時(shí)期瞬時(shí)表達(dá)。【結(jié)論】豬NR4A1基因的CDS區(qū)在物種間保守性較強(qiáng),推測(cè)豬NR4A1基因調(diào)節(jié)卵泡的發(fā)育和排卵過(guò)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.
【作者單位】： 安徽省農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所;華中農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)院;
【基金】：安徽省農(nóng)業(yè)科學(xué)院科技創(chuàng)新團(tuán)隊(duì)項(xiàng)目(13C0405);安徽省農(nóng)業(yè)科學(xué)院院長(zhǎng)青年基金項(xiàng)目(14B0427)
【分類(lèi)號(hào)】：S828
【正文快照】： NR4A1基因作為一個(gè)孤核受體的轉(zhuǎn)錄因子,編碼類(lèi)固醇-甲狀腺激素超家族。NR4A1蛋白是一類(lèi)配體依賴轉(zhuǎn)錄調(diào)節(jié)蛋白[1],它在轉(zhuǎn)錄過(guò)程中起作用的是N-末端,參與調(diào)節(jié)核定位和轉(zhuǎn)錄激活過(guò)程的是C-末端[2-4]。在細(xì)胞中進(jìn)行的類(lèi)固醇合成活性研究結(jié)果顯示,NR4A1基因表達(dá)與P450膽固醇側(cè)鏈裂解

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