鴨Ccna1啟動(dòng)子克
發(fā)布時(shí)間：2018-10-21 17:27
【摘要】：為研究Ccna1基因在鴨骨骼肌發(fā)育中的作用,本研究應(yīng)用RT-PCR擴(kuò)增和克隆了鴨Ccna1基因啟動(dòng)子區(qū)并作生物信息學(xué)分析,應(yīng)用熒光定量PCR技術(shù)檢測(cè)Ccna1基因以及參與該基因表達(dá)調(diào)控的多個(gè)肌源性轉(zhuǎn)錄因子在鴨胚骨骼肌組織發(fā)育過程中的表達(dá)模式。結(jié)果表明,擴(kuò)增得到鴨Ccna1基因啟動(dòng)子2 213bp。序列分析表明,鴨Ccna1啟動(dòng)子存在典型的TATA-box、CAAT-box調(diào)控元件,有MyoD、MRF4、MyoG、Sp1、PITX1及MEF2等多個(gè)轉(zhuǎn)錄因子結(jié)合位點(diǎn)。定量結(jié)果發(fā)現(xiàn),MyoD、MRF4、MyoG和Ccna1在鴨胚胎骨骼肌發(fā)育過程中均有表達(dá)。在鴨胸肌中,MRF4在E23時(shí)期表達(dá)量最高,顯著高于D8的表達(dá)量(P0.05);在腿肌中,MRF4和MyoG表達(dá)量在E17時(shí)期達(dá)到最高,且顯著高于D2、D8(P0.05)。聚類分析表明,在鴨胸肌組織中Ccna1表達(dá)模式與MyoD一致,在腿肌組織中Ccna1表達(dá)模式與MRF4、MyoG一致。初步推測(cè),Ccna1參與到了鴨骨骼肌組織的發(fā)育,且在鴨腿肌的生長(zhǎng)發(fā)育中,MRF4、MyoG可能與Ccna1相互作用進(jìn)而調(diào)控骨骼肌發(fā)育,而在鴨胸肌組織中,Ccna1的表達(dá)可能與MyoD的轉(zhuǎn)錄調(diào)控有關(guān)。
[Abstract]:In order to study the role of Ccna1 gene in the development of duck skeletal muscle, the promoter region of duck Ccna1 gene was amplified and cloned by RT-PCR and analyzed by bioinformatics. Fluorescence quantitative PCR was used to detect the expression patterns of Ccna1 gene and several myogenic transcription factors involved in the regulation of the gene expression in the skeletal muscle of duck embryo. The results showed that the promoter of duck Ccna1 gene was 2213bp. Sequence analysis showed that there were typical TATA-box,CAAT-box regulatory elements in duck Ccna1 promoter and several transcription factor binding sites such as MyoD,MRF4,MyoG,Sp1,PITX1 and MEF2. The quantitative results showed that both MyoD,MRF4,MyoG and Ccna1 were expressed during the development of duck embryonic skeletal muscle. In Duck pectoralis muscle, the expression of MRF4 was the highest in E23 stage, significantly higher than that of D8 (P0.05); in leg muscle, the expression of MRF4 and MyoG reached the highest level at E17 stage, and significantly higher than that of D2D8 (P0.05). Cluster analysis showed that the expression pattern of Ccna1 was the same as that of MyoD in the breast muscle of duck, and that of Ccna1 in the leg muscle was the same as that in the tissue of the leg muscle. It is suggested that Ccna1 is involved in the development of duck skeletal muscle, and MRF4,MyoG may interact with Ccna1 to regulate skeletal muscle development during the growth and development of duck leg muscle, while in duck breast muscle, the expression of Ccna1 may be related to the transcriptional regulation of MyoD.
【作者單位】： 四川農(nóng)業(yè)大學(xué)動(dòng)物遺傳育種研究所;
【基金】：國家自然科學(xué)基金(31301964) 大學(xué)生創(chuàng)新性實(shí)驗(yàn)項(xiàng)目(1510626040)
【分類號(hào)】：S834
本文編號(hào)：2285826
[Abstract]:In order to study the role of Ccna1 gene in the development of duck skeletal muscle, the promoter region of duck Ccna1 gene was amplified and cloned by RT-PCR and analyzed by bioinformatics. Fluorescence quantitative PCR was used to detect the expression patterns of Ccna1 gene and several myogenic transcription factors involved in the regulation of the gene expression in the skeletal muscle of duck embryo. The results showed that the promoter of duck Ccna1 gene was 2213bp. Sequence analysis showed that there were typical TATA-box,CAAT-box regulatory elements in duck Ccna1 promoter and several transcription factor binding sites such as MyoD,MRF4,MyoG,Sp1,PITX1 and MEF2. The quantitative results showed that both MyoD,MRF4,MyoG and Ccna1 were expressed during the development of duck embryonic skeletal muscle. In Duck pectoralis muscle, the expression of MRF4 was the highest in E23 stage, significantly higher than that of D8 (P0.05); in leg muscle, the expression of MRF4 and MyoG reached the highest level at E17 stage, and significantly higher than that of D2D8 (P0.05). Cluster analysis showed that the expression pattern of Ccna1 was the same as that of MyoD in the breast muscle of duck, and that of Ccna1 in the leg muscle was the same as that in the tissue of the leg muscle. It is suggested that Ccna1 is involved in the development of duck skeletal muscle, and MRF4,MyoG may interact with Ccna1 to regulate skeletal muscle development during the growth and development of duck leg muscle, while in duck breast muscle, the expression of Ccna1 may be related to the transcriptional regulation of MyoD.
【作者單位】： 四川農(nóng)業(yè)大學(xué)動(dòng)物遺傳育種研究所;
【基金】：國家自然科學(xué)基金(31301964) 大學(xué)生創(chuàng)新性實(shí)驗(yàn)項(xiàng)目(1510626040)
【分類號(hào)】：S834
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1 宋美華;乳腺癌組織CCNA1啟動(dòng)子區(qū)甲基化與腫瘤轉(zhuǎn)移的相關(guān)性[D];南京醫(yī)科大學(xué);2011年
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