【摘要】 本研究旨在探讨YWHAG(Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein,gamma polypeptide)基因在猪卵泡发育中的表达规律及其与卵泡发育的关系。从猪卵泡组织中克隆YWHAG基因CDS(Coding region)区全序列,采用半定量RT-PCR和Real-time PCR方法检测YWHAG基因在不同组织中的表达谱信息,并进一步分析了该基因在梅山、杜洛克猪L、M2、M1、S各级别卵泡中的表达变化。结果显示:克隆得到了猪YWHAG基因780bp编码区全序列,通过与人、小鼠、牛YWHAG基因CDS区BLAST比对,其同源性分别为97.72%、96.68%和98.57%。YWHAG基因在肌肉、肝、脂肪、肺、心、脾、肾、子宫、输卵管、小肠、胃、卵巢、垂体、下丘脑等组织中均有表达,尤其在脂肪、小肠和大脑中表达较高。YWHAG基因在不同直径卵泡中的表达量的趋势均为M1卵泡和L卵泡最高,M2卵泡次之,S卵泡最低,并且在M1卵泡和L卵泡中的表达量显著高于M2卵泡和S卵泡(P<0.05)。YWHAG基因在梅山猪各级别卵泡中的表达量均高于杜洛克猪(P<0.05),并且在梅山猪S和M2卵泡的表达量分别极显著高于杜洛克猪(P<0.01),而在M1卵泡及L卵泡中的表达量无显著差异;结果提示,YWHAG基因可能参与猪卵泡发育中的凋亡过程,从而引起梅山与杜洛克猪在排卵数方面存在差异。更多还原

【Abstract】 To detect the expression pattern of YWHAG(Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein,gamma polypeptide)gene in during porcine follicle development.The CDS(Coding sequence)of YWHAG gene was cloned from porcine follicular tissue. Semi-quantitative RT-PCR was used to analyze the tissue expression profiling of YWHAGgenes, Furthermore the expression level of YWHAGin follicles with different diameter(L,M2,M1,S) from Duroc and Meishan were investigated by real-time fluorescence quantitative PCR.The results showed that:the length of CDS of porcine YWHAG was 780bp,and it’s similarity with that of human,mouse,and cattle was 97.72%,96.68% and 98.57%,respectively.YWHAG mRNA was expressed in all tested tissues(Muscle,fat,heart,liver,spleen,lung,kidney,stomach,small intestine,ovarian,uterine,uterine horn,tubal,pituitary,corpus luteum,brain,the hypothalamus), and especially high in fat,intestine and brain.Both in Duroc and Meishan swine,the expression of YWHAG mRNA in M1and L follicles were significantly higher than M2and S follicles(P<0.05).Moreover,The expression in S and M2follicles from Meishan were significantly higher than that from Duroc(P<0.01),while no significant difference in M1and L follicle.In conclusion,YWHAGgene may be involved in the apoptosis of porcine follicular development process,resulting in differences of ovulation rate in swine.更多还原