滩羊皮肤组织差异蛋白质组学研究

发布时间：2018-05-02 00:47

本文选题：滩羊 + 皮肤组织　；参考：《宁夏大学》2017年硕士论文

【摘要】：滩羊是我国唯一生产名贵二毛裘皮的特有绵羊品种,因其特殊地理生态环境的长期自然选择和人工选育,造就了宁夏滩羊集产肉、产毛、产皮为一体的名贵羊种。滩羊裘皮以美观、轻便、保暖、耐用而闻名,其在二毛期由于毛股花形不同,因而表现出不同的花穗类型,但随着二毛期滩羊的日龄增加,其裘皮毛股弯曲等特征会逐渐减弱直至消失。滩羊皮肤作为这些特征的承载者,通过对皮肤蛋白质组的研究,从而了解其内在原因。基于上述背景,本研究开展以下方面的工作:利用双向凝胶电泳(2-DE)技术研究一周岁内滩羊皮肤差异表达蛋白,对一周岁内4个生长时期的滩羊皮肤蛋白图谱进行两两之间进行比对,共发现了 19个差异蛋白点,质谱成功鉴定出15个蛋白点共13种蛋白。可能与滩羊皮肤生长调控有关的蛋白有14-3-3蛋白σ亚型、膜联蛋白A2、角蛋白25、微管蛋白α链。对差异表达蛋白进行分析,发现这些差异表达蛋白可能与滩羊皮肤不同生长阶段的毛囊呈周期性变化有关。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、中卫山羊和新吉细毛羊的羊皮肤蛋白进行差异表达研究,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;最后对差异蛋白进行GO和Pathway分析。四种羊皮肤中共鉴定到1161个蛋白,其中内蒙古苏尼特羊与滩羊比对中发现30个差异蛋白,中卫山羊与滩羊比对发现112个差异蛋白,新吉细毛羊比对与滩羊发现107个差异蛋白,对各比对组的皮肤差异蛋白进行分析,发现发现KRTAP6、KRTAP11.1、KRT25、KRT34、KRT83和TCHH可能与滩羊羊毛粗细及弯曲度形成相关。应用iTRAQ蛋白质组学方法,联合LC-MS/MS技术对滩羊二毛期串子花型、软大花型、绿豆丝型和其它不规则型4种花穗型裘皮蛋白进行差异表达研究,在4种花穗型裘皮共鉴定得2886个蛋白,软大花型与串子花型比对获得差异蛋白数为135个;绿豆丝型与串子花型比对获得差异蛋白142个;不规则型与串子花型比对获得差异蛋白为113个,对表达差异蛋白进行分析,发现膜联蛋白与VEGF信号通路与软大花型毛股形成相关,KRTAP3和KRTAP6可能与滩羊串子花型毛股结构相关。上述研究为滩羊皮肤的年变化规律和优质二毛裘皮选育以及滩羊串子花型二毛裘皮育种提供理论基础和科学依据。
[Abstract]:Tan sheep is the only unique sheep breed producing rare and rare fur in China. Due to the long-term natural selection and artificial breeding of its special geographical and ecological environment, Tan sheep in Ningxia is a rare sheep that produces meat, wool and skin. Tan Sheep fur is famous for its beauty, lightness, warmth, durability and durability. It shows different flower spike types in the period of 20 wool because of the different flower shape, but with the increase of the age of Tan sheep in the period of 20 wool, Its fur hair and other characteristics such as thigh bending will gradually weaken until disappear. As the carrier of these characteristics, the skin of Tan Sheep is studied on the proteome of skin to understand its internal causes. Based on the above background, this study carried out the following work: Two-Dimensional Gel Electrophoresis (2-DEE) technique was used to study the differentially expressed proteins in the skin of Tan Sheep within one year old. A total of 19 differential protein spots were found in the skin protein maps of Tan sheep in four growing stages within one year of age, and 13 proteins were successfully identified by mass spectrometry. There are 14-3-3 protein 蟽 subtypes, annexin A2, keratin 25 and tubulin 伪 chain, which may be related to the regulation of skin growth in Tan sheep. It was found that these differentially expressed proteins might be related to the periodic changes of hair follicles in different growth stages of Tan sheep skin. The differential expression of sheep skin protein in Tan sheep, Inner Mongolia Sunita, Zhongwei goat and Xinji fine wool sheep was studied by using isotopic labeling relative and absolute quantitative ITRAQ proteomics combined with Multidimensional liquid Chromatography tandem Mass Spectrometry (MLDMS) technique in Tan sheep, Inner Mongolia Sunita, Zhongwei goat and Xinji fine wool sheep. The differential protein was identified by using Proteome Discoverer 1.4 software and database search. Finally, the differential protein was analyzed by go and Pathway. A total of 1,161 proteins were identified in the skin of four kinds of sheep, of which 30 were found in the comparison of Sunita sheep and Tan sheep in Inner Mongolia, 112 differential proteins in Zhongwei goat and Tan sheep, 107 differential proteins in Xinji fine wool sheep and Tan sheep. By analyzing the skin differential proteins of the control groups, it was found that KRTAP6, KRTAP11.1, KRT25, KRT34, KRT83 and TCHH might be related to the formation of wool thickness and curvature of Tan sheep. ITRAQ proteomics and LC-MS/MS technique were used to study the differential expression of Fur skin proteins in four flower spike types of Tan Sheep, such as flower pattern, soft large flower type, mung bean silk type and other irregular type. A total of 2886 proteins were identified in four flower panicle types of fur. The number of differentially expressed proteins was 135 in the ratio of soft large flower type to string flower type, 142 in mung bean silk and string flower type. 113 differentially expressed proteins were obtained by comparison of irregular and string flower types. It was found that VEGF signal pathway and VEGF signal pathway were related to the formation of soft large floral hair strands. KRTAP3 and KRTAP6 might be related to the floral pattern of hair strands of Tan sheep. The above studies provide theoretical and scientific basis for the annual variation of Tan sheep skin and the breeding of fine quality Ermao fur and the breeding of Tan sheep's flower pattern of Ermao fur.
【学位授予单位】：宁夏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S826

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