本文選題：鴨 + Pax3　； 參考：《四川農(nóng)業(yè)大學》2015年碩士論文

【摘要】：衛(wèi)星細胞作為肌肉胚后發(fā)育的干細胞,會直接影響肌肉的生長發(fā)育。Pax3和Pax7是衛(wèi)星細胞的標志性基因,Pax3對于胚胎期動物的四肢正常地生長發(fā)育必不可少,而Pax7對肌肉的損傷修復有重要作用。它們對于肌肉的生長發(fā)育都很重要。農(nóng)華肉鴨P系、建昌鴨、黑鶩鴨三個鴨品種在產(chǎn)肉性能上具有很大的差異,為了探討Pax3及相關(guān)基因與這幾個品種產(chǎn)肉性能間的關(guān)系,本研究通過擴增Pax3啟動子序列并進行直接測序,篩選Pax3基因啟動子區(qū)域是否存在影響產(chǎn)肉性能的SNP位點。同時,選擇Pax3、Pax7、MyoD、MyoG為候選基因,檢測它們在北京鴨、建昌鴨和黑鶩鴨三個品種腿肌中的表達差異。結(jié)果表明：1.擴增出鴨Pax3基因上游1271bp的啟動子序列。生物信息學分析結(jié)果表明,鴨Pax3啟動子區(qū)內(nèi)存在典型的CAAT-Box、TATA-Box、G-Box、GATA-Box等結(jié)構(gòu)域以及GATA-1、Sp-1和Myod轉(zhuǎn)錄結(jié)合位點,無CpG島。2.通過對Pax3啟動子片段的測序,共發(fā)現(xiàn)26個可能的SNP位點,但僅有5個SNP位點符合哈迪-溫伯格定律(SNP1、SNP16、SNP19、SNP21和SNP24)。各SNP與三個鴨品種屠體性狀的相關(guān)分析結(jié)果顯示,農(nóng)華肉鴨P系和黑鶩鴨中SNP1和SNP16的BB型個體的腹脂重、皮脂重和腿脂重高于AB型；在黑鶩鴨品種中SNP19的BB型個體腹脂重、皮脂重、腿脂重、腿肌重和胸肌重都高于AB型；建昌鴨和黑鶩鴨中SNP21的AB型個體腹脂重、皮脂重、腿脂重都高于BB型；建昌鴨SNP 24的AB型個體的腹脂重、皮脂重、腿脂重高于BB型,但以上差異均未達顯著水平(P0.05)。3.農(nóng)華肉鴨P系腿肌中Pax3和Pax7的基因表達量顯著高于建昌鴨和黑鶩鴨(P0.05)；MyoD基因表達量在黑鶩鴨腿肌中最高,建昌鴨和農(nóng)華肉鴨P系依次降低,但差異不顯著(P0.05)；MyoG基因在建昌鴨腿肌中的表達量顯著高于農(nóng)華肉鴨P系和黑鶩鴨(P0.05)。不同品種腿肌中MyoG的表達量與腿肌重間不存在線性關(guān)系,表明MyoG的表達可能與肌肉的沉積無顯著相關(guān)。4.在蛋白表達水平,Pax3和Pax7有相同的趨勢,均是在農(nóng)華肉鴨P系腿肌中最高,在黑鶩鴨腿肌中最低,但是差異不顯著(P0.05)。5.農(nóng)華肉鴨P系腿肌中Pax3和Pax7在mRNA和蛋白水平的表達量均最高,表明Pax3和Pax7的表達可能是農(nóng)華肉鴨P系相對建昌鴨和黑鶩鴨具有更高產(chǎn)肉量的原因之一。
[Abstract]:Satellite cells, as stem cells for post-embryonic muscle development, directly affect muscle growth and development. Pax3 and Pax7 are the iconic genes of satellite cells that are essential for the normal growth and development of the limbs of embryonic animals.Pax7 plays an important role in the repair of muscle injury.They are important for the growth and development of muscles.In order to explore the relationship between Pax3 and related genes, there are great differences in meat production performance among three breeds of Nonghua Meat Duck, Jianchang Duck and Black Duck, in order to explore the relationship between them.In this study, the Pax3 promoter sequence was amplified and sequenced directly to screen the presence of SNP sites in the promoter region of the Pax3 gene.At the same time, Pax3, Pax7 and MyoDG were selected as candidate genes to detect their expression differences in leg muscles of Beijing duck, Jianchang duck and black duck.The result shows that 1: 1.The promoter sequence of 1271bp in upstream of duck Pax3 gene was amplified.Bioinformatics analysis showed that there were typical CAAT-BoxTa-TA-BoxTa-G-BoxTa-Box domains and GATA-1Sp-1 and Myod transcriptional binding sites in duck Pax3 promoter region, and there was no CpG island. 2.By sequencing the Pax3 promoter fragments, 26 possible SNP loci were found, but only 5 SNP loci were in accordance with the Hardy Weinberg's Law: SNP1, SNP16, SNP19, SNP21 and SNP24.The results of correlation analysis between SNP and carcass traits of three duck breeds showed that the abdominal fat weight, sebum weight and leg fat weight of BB individuals of SNP1 and SNP16 in P line and black duck of Nong Hua duck were higher than those of AB type, and the abdominal fat weight of BB type individuals of SNP19 in black duck breeds was higher than that of AB type.Sebum weight, leg fat weight, leg muscle weight and breast muscle weight were all higher than AB type; AB type individual abdominal fat weight, sebum weight and leg fat weight of SNP21 in Jianchang duck and black duck were all higher than BB type; and SNP 24 AB individual had abdominal fat weight and sebum weight in Jianchang duck.The leg fat weight was higher than that of BB type, but the difference was not significant (P 0.05. 3).The expression of Pax3 and Pax7 in the leg muscle of P line of Nonghua duck was significantly higher than that of Jianchang duck and black duck. The expression of Pax3 and Pax7 was the highest in the leg muscle of black duck, and the P line of Jianchang duck and Nong Hua duck decreased in turn.However, the expression of MyoG gene in leg muscle of Jianchang duck was significantly higher than that of P line of Nong Hua Meat Duck and P0.05 of Black Duck.There was no linear relationship between the expression of MyoG and leg muscle weight in different leg muscles, indicating that the expression of MyoG may not be significantly related to muscle deposition.The protein expression level of Pax3 was the same as that of Pax7, which was the highest in the P leg muscle of Nonghua duck and the lowest in the leg muscle of black duck, but there was no significant difference between them.The expression of Pax3 and Pax7 in the leg muscle of Nonghua duck was the highest in mRNA and protein levels, which suggested that the expression of Pax3 and Pax7 might be one of the reasons for the higher meat yield of P line compared with Jianchang duck and black duck.
【學位授予單位】：四川農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S834

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