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多房棘球绦虫原头节蛋白质组学研究
时间:2011-01-23 浏览次数:1586次 无忧论文网
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动物学
寄生虫蛋白质组学寄生虫蛋白质组学
    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。 [英文摘要]:     The proteins of Europe Echinococcus multilocularis protoscolece were analyzed by two-dimension electrophoresis (2-DE) separating technique, Western-blotting, Mass spectrometry (MS) technique, and protein bioinformatics. After analyzed 2-DE pattern by ImageMaster software, about 485±18 protein spots were acquired, their molecular weight were about 10-120kDa and the isoelectric points were about 3.5-9. Then 90 protein spots rich in content were chosen and identified by MS to acquire their Peptide Mass Fingerprint(PMF). After analyzed by the software of Mascot, some kinds of proteins highly related to the growth ,development, movement and regulation of the parasite were aquired, which included: actin, transketolase, thioredoxin peroxidase, glutathione transferase, HSP70, HSP20, actin-binding and severin family group-like protein, paramyosin, microtubulin, 78kD glucose regulated protein, tropomyosin, 14-3-3 protein, Caspase-3, hydatid disease diagnostic antigen P-29, and etc. Result from the analysis of antigens expression of Europe Echinococcus multilocularis protoscolece showed, cytoskeleton proteins and chaperonins are the main antigens referring to the host—parasite interaction. Further studies on this field will not only offer some clues about the host—parasite interaction mechanism, but all these relative reseaches will become the bases of finding efficient vaccine candidates and drug targets.
    What’s more, the protein and antigen patterns of Europe Echinococcus multilocularis were also contrasted with those of Echinococcus sibiricensis. As a result, there were only minor differences in protein expression level, but some obvious differences between the antigen patterns of these two types of Echinococcus are found, for example, there is a differential 124kDa antigenic band in Europe Echinococcus multilocularis. While the HSP70 and paramyosin antigens of Echinococcus sibiricensis induce stronger immune response than the former. All these results show that the two types of echinococcus are immunologically different.    
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