多房棘球绦虫原头节蛋白质组学研究

发布时间：2018-12-07 20:02

【摘要】： 本文运用蛋白质组学中的双向电泳分离技术、质谱分析技术和蛋白质信息学方法及免疫蛋白质组学中的Western-blotting技术主要对欧洲多房棘球绦虫原头节蛋白及抗原表达谱进行研究。利用ImageMaster软件分析欧洲多房棘球绦虫原头节的双向电泳图谱,共分离到485±18个蛋白斑点,这些蛋白质斑点大部分分子量介于10-120kD、等电点3.5-9的区域。挑选出其中丰度较高的蛋白斑点进行质谱鉴定,结果鉴定出一些与寄生虫生长、发育、运动、调控相关的蛋白,包括钙网蛋白、微管蛋白、转醇酮酶、硫氧还过氧化物酶、谷胱甘肽S转移酶、HSP90、HSP20、HSP70、肌动蛋白结合与切割蛋白(actin-binding and severin family group-like protein)、副肌球蛋白、肌动蛋白、肌球蛋白样蛋白(myosin-like protein)、78KD葡萄糖调节蛋白、胱冬酶3、14-3-3蛋白、原肌球蛋白、包虫病诊断性抗原P-29等。分析其抗原蛋白表达谱发现,参与寄生虫-宿主相互作用的抗原主要是细胞骨架类蛋白及分子伴侣类蛋白。深入研究这些主要抗原成分,将有利于阐明寄生虫-宿主相互作用机制,并且为寻找有效的疫苗候选分子及药物作用靶标奠定基础。此外,本文还比较了欧洲多房棘球绦虫原头节与西伯利亚棘球绦虫原头节蛋白表达谱、抗原蛋白谱之间的差异,结果发现两者之间在总体蛋白表达上差异不明显,但是它们的抗原蛋白谱之间确有较明显的差异,如欧洲多房棘球绦虫在约120kDa处出现特异的免疫印迹条带,鉴定结果表明,该处蛋白可能为类似于海胆的转座酶,而西伯利亚棘球绦虫在副肌球蛋白、HSP70等处的抗原抗体反应明显强于欧洲多房棘球绦虫。这一结果表明,两虫种在免疫学上存在差异
[Abstract]:In this paper, two dimensional electrophoretic separation of Echinococcus multilocularis, mass spectrometry, protein informatics and Western-blotting technique in immuno proteomics were used to study the expression profiles of protoganglion proteins and antigens of Echinococcus multilocularis (Echinococcus multilocularis). A total of 485 卤18 protein spots were isolated from Echinococcus multilocularis by using ImageMaster software. The molecular weight of most of these spots ranged from 10 to 120 KD and the isoelectric point was 3.5-9. Some proteins related to the growth, development, movement and regulation of the parasite were identified, including calcium reticin, tubulin, transketonase, thioredoxidase, and thioredoxidase. Glutathione S-transferase, HSP90,HSP20,HSP70, actin binding and cleavage protein (actin-binding and severin family group-like protein), accessory myosin, actin, myosin-like protein), 78KD glucose-regulated protein) Cysteinase 314-3-3 protein, promyosin, hydatidosis diagnostic antigen P-29, etc. It was found that cytoskeleton proteins and molecular chaperones were the main antigens involved in parasita-host interaction. Further study of these major antigens will help to clarify the mechanism of parasite-host interaction and lay a foundation for finding effective vaccine candidate molecules and drug action targets. In addition, the differences of protein expression profiles and antigenic protein profiles between Echinococcus multilocularis and Echinococcus Siberia were compared. The results showed that there was no significant difference in total protein expression between Echinococcus multilocularis and Echinococcus Siberia. However, there are obvious differences between their antigenic protein profiles. For example, Echinococcus multilocularis has a specific Western blotting band at about 120kDa, which may be a transpose-like enzyme similar to sea urchin. The antigen-antibody responses of Echinococcus Siberia were stronger than those of Echinococcus multilocularis in Europe. The results show that there are immunological differences between the two species.
【学位授予单位】：厦门大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R383;S852.7

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