Pax3基因启动子区SNP及相关基因表达与鸭产肉性能的关系研究

发布时间：2018-04-16 00:24

  本文选题：鸭 + Pax3　； 参考：《四川农业大学》2015年硕士论文

【摘要】：卫星细胞作为肌肉胚后发育的干细胞,会直接影响肌肉的生长发育。Pax3和Pax7是卫星细胞的标志性基因,Pax3对于胚胎期动物的四肢正常地生长发育必不可少,而Pax7对肌肉的损伤修复有重要作用。它们对于肌肉的生长发育都很重要。农华肉鸭P系、建昌鸭、黑鹜鸭三个鸭品种在产肉性能上具有很大的差异,为了探讨Pax3及相关基因与这几个品种产肉性能间的关系,本研究通过扩增Pax3启动子序列并进行直接测序,筛选Pax3基因启动子区域是否存在影响产肉性能的SNP位点。同时,选择Pax3、Pax7、MyoD、MyoG为候选基因,检测它们在北京鸭、建昌鸭和黑鹜鸭三个品种腿肌中的表达差异。结果表明：1.扩增出鸭Pax3基因上游1271bp的启动子序列。生物信息学分析结果表明,鸭Pax3启动子区内存在典型的CAAT-Box、TATA-Box、G-Box、GATA-Box等结构域以及GATA-1、Sp-1和Myod转录结合位点,无CpG岛。2.通过对Pax3启动子片段的测序,共发现26个可能的SNP位点,但仅有5个SNP位点符合哈迪-温伯格定律(SNP1、SNP16、SNP19、SNP21和SNP24)。各SNP与三个鸭品种屠体性状的相关分析结果显示,农华肉鸭P系和黑鹜鸭中SNP1和SNP16的BB型个体的腹脂重、皮脂重和腿脂重高于AB型；在黑鹜鸭品种中SNP19的BB型个体腹脂重、皮脂重、腿脂重、腿肌重和胸肌重都高于AB型；建昌鸭和黑鹜鸭中SNP21的AB型个体腹脂重、皮脂重、腿脂重都高于BB型；建昌鸭SNP 24的AB型个体的腹脂重、皮脂重、腿脂重高于BB型,但以上差异均未达显著水平(P0.05)。3.农华肉鸭P系腿肌中Pax3和Pax7的基因表达量显著高于建昌鸭和黑鹜鸭(P0.05)；MyoD基因表达量在黑鹜鸭腿肌中最高,建昌鸭和农华肉鸭P系依次降低,但差异不显著(P0.05)；MyoG基因在建昌鸭腿肌中的表达量显著高于农华肉鸭P系和黑鹜鸭(P0.05)。不同品种腿肌中MyoG的表达量与腿肌重间不存在线性关系,表明MyoG的表达可能与肌肉的沉积无显著相关。4.在蛋白表达水平,Pax3和Pax7有相同的趋势,均是在农华肉鸭P系腿肌中最高,在黑鹜鸭腿肌中最低,但是差异不显著(P0.05)。5.农华肉鸭P系腿肌中Pax3和Pax7在mRNA和蛋白水平的表达量均最高,表明Pax3和Pax7的表达可能是农华肉鸭P系相对建昌鸭和黑鹜鸭具有更高产肉量的原因之一。
[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.
【学位授予单位】：四川农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S834

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