【摘要】：目的:研究CBR1基因啟動子在豬子宮內(nèi)膜細(xì)胞的表達(dá)調(diào)控機(jī)制。創(chuàng)新點(diǎn):發(fā)現(xiàn)CBR1基因啟動子在豬子宮內(nèi)膜受到炎性因子核轉(zhuǎn)錄因子kappa B(NF-κB)成員p65調(diào)控。p65對該啟動子具有正向調(diào)節(jié)作用,但是對于CBR1基因的表達(dá)并不是必需的。方法:通過雙熒光素酶報告基因載體確定CBR1基因啟動子轉(zhuǎn)錄活性區(qū),通過染色質(zhì)免疫沉淀(Ch IP)技術(shù)確定p65能夠結(jié)合CBR1基因啟動子,通過超表達(dá)和干擾表達(dá)實(shí)驗(yàn)證實(shí)p65對CBR1基因啟動子的調(diào)控作用。結(jié)論:豬CBR1基因啟動子-1640/-647區(qū)對于其轉(zhuǎn)錄活性是必需的,在-1545/-1531區(qū)存在p65的結(jié)合位點(diǎn)。p65在豬子宮內(nèi)膜細(xì)胞中促進(jìn)了CBR1基因mRNA的表達(dá),但是干擾p65則不會造成CBR1基因mRNA表達(dá)量下降,推斷p65不是CBR1基因表達(dá)的必需因素。
[Abstract]:Aim: to study the expression and regulation mechanism of CBR1 gene promoter in porcine endometrial cells. Innovation: it is found that the promoter of CBR1 gene is regulated by p65 member of kappa B (NF- kappa B in porcine endometrium. P65 has a positive regulatory effect on the promoter, but it is not necessary for the expression of CBR1 gene. Methods: the transcriptional active region of CBR1 gene promoter was determined by double luciferase reporter gene vector, and p65 could bind to CBR1 gene promoter by chromatin immunoprecipitation (Ch IP) technique. The regulatory effect of p65 on the promoter of CBR1 gene was confirmed by over-expression and interference expression experiments. Conclusion: the transcriptional activity of porcine CBR1 gene promoter-1640 is necessary, and p65 binding site exists in-1545 / 1531 region. P65 promotes the expression of CBR1 gene mRNA in porcine endometrial cells. However, interference with p65 did not result in the decrease of mRNA expression of CBR1 gene, and it was inferred that p65 was not an essential factor in the expression of CBR1 gene.
【作者單位】： Guangdong
【基金】：Project supported by the National Natural Science Foundation of China(No.31201771) the Earmarked Fund for China Agriculture Research System(No.CARS-36)
【分類號】：S828

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