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猪CCT5基因的cDNA克隆、序列分析及时空表达

DOI: 10.7685/j.issn.1000-2030.2013.04.016, PP. 99-104

Keywords: ,CCT5,骨骼肌,克隆,表达

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Abstract:

基于2-DE和MS的蛋白质组学方法分析梅山猪骨骼肌发育过程中蛋白质动态表达变化,发现CCT5差异表达。为进一步获得猪CCT5基因的序列特征、时空表达及生物学功能等相关信息,克隆猪CCT5基因,分析不同物种序列进化关系,检测CCT5在猪不同组织和骨骼肌不同发育阶段中的表达。序列分析结果表明在获得的2286bp猪CCT5cDNA序列中包括1626bp的完整开放阅读框(ORF),编码541个氨基酸,其氨基酸序列与人的相似性为97%,表明该蛋白在不同物种间高度保守。荧光定量PCR结果显示,CCT5基因在猪肌肉和脂肪中表达量最高,心、肝等组织中表达量很低,没有肌纤维特异性;不同发育阶段骨骼肌中,该基因在出生后3d的表达量最高,胚胎期表达量最低;在不同品种间,胚胎期及出生后3、30和180d的表达量大白猪显著高于梅山猪(P<0.05),而60d却显著低于梅山猪(P<0.05)。结果提示CCT5基因在猪骨骼肌发育过程中起着重要作用。

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