应用iTRAQ技术对滩羊裘皮差异表达蛋白的研究

Study on Differentially Expressed Proteins of Tan Sheep Skin Using iTRAQ Technology

为了研究与滩羊羊毛纤维粗细及弯曲度和特定花穗构成相关的差异蛋白,试验应用同位素标记相对和绝对定量(iTRAQ)蛋白质组学方法,联合多维液相色谱串联质谱(LC-MS/MS)技术对滩羊、内蒙古苏尼特羊、中卫山羊和新吉细毛羊的羊皮肤蛋白进行鉴定和筛选,并应用Proteome Discoverer 1.4软件进行定量分析,结合数据库搜索,鉴定出差异蛋白;最后应用生物信息学技术对差异蛋白进行GO分析和Pathway分析。4种羊皮肤中共鉴定到1 161个蛋白,其中内蒙古苏尼特羊与滩羊比对发现30个差异蛋白,中卫山羊与滩羊比对发现112个差异蛋白,新吉细毛羊与滩羊比对发现107个差异蛋白,对各比对组的皮肤差异蛋白进行分析,发现14-3-3蛋白σ亚型、KRT25、KRT34、KRT85、TCHH、KAP6和KAP11.1可能与滩羊羊毛纤维粗细及弯曲度和特定花穗形状的形成有关。本研究可为滩羊优质二毛裘皮选育研究提供科学依据。

In order to study the differential protein associated with the thickness and curvature of the flax fiber and the specific spike composition of Tan sheep,this study used the isotope labeling relative and absolute quantification（iTRAQ）proteomics methods,and combined with multidimensional liquid chromatography tandem mass spectrometry（LC-MS/MS）identification and screening of skin protein from Tan sheep,Inner Mongolia Sunitou sheep,Zhongwei goat and Xinji Fine-wool sheep,and the Proteome Discoverer 1.4software was used for quantitative analysis,combined with database search,identification of differential protein.Finally,the differential proteins was analyzed by the GO and Pathway analysis methods.1 161 proteins were identified from the four kinds of sheep skin,30 different proteins were found between Inner Mongolia Sunitou sheep and Tan sheep,112 differential proteins were found between Zhongwei goats and Tan sheep,107 differential proteins were found between Xinji Fine-wool sheep and Tan sheep,the skin differential proteins were analyzed for each group,it was found that the 14-3-3protein sigma,KRT25,KRT34,KRT85,TCHH,KAP6 and KAP11.1 might be related to the thickness and curvature of the flax fiber and the formation of specific spike shape of Tan sheep.The result would provide a scientific basis for the breeding of Tan sheep fur quality.