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研究生:彭介甫
研究生(外文):Chieh-Fu Peng
論文名稱:花笠螺與細線青螺齒舌礦化的蛋白質體分析
論文名稱(外文):Proteome Analysis of Biomineralization of Radular Teeth in the Limpets Cellana Toreuma and Patelloida striata
指導教授:李家維李家維引用關係
指導教授(外文):Chia-Wei Li
學位類別:碩士
校院名稱:國立清華大學
系所名稱:生命科學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:英文
論文頁數:122
中文關鍵詞:生物礦化齒舌蛋白質體分析花笠螺細線青螺
外文關鍵詞:BiomineralizationRadularProteome AnalysisLimpetCellana ToreumaPatelloida striata
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齒舌是軟體動物的攝食器。笠螺及青螺的齒舌在不同的發育階段會相繼累積鐵和矽元素,是研究生物礦化極佳的材料。齒舌的發育及礦化是一連續且同時性的事件,亦即可以在一條齒舌上可關察到一個攝食器官的整個發育過程。我們利用蛋白質體的方法分析參與細線青螺與花笠螺齒舌不同發育階段生物礦化的蛋白質。在不同發育階段,S35標記的蛋白質在SDS-PAGE上顯示了不同的模式,指出齒舌發育會經過某些變化,而這些變化可能與齒舌的生物礦化過程有關。有新合成的蛋白質發現被運送至細胞外的齒舌牙腔內。花笠螺和細線青螺在二維蛋白質電泳中分別出現了一千及三百個點,比較齒舌的不同發育階段後發現,分別有17及19個蛋白質在齒舌形成的不同發育階段有差異性的表現。例如我們發現在花笠螺中,一系列蛋白質(點17、20、21、22、23、24、25)和點53只有在齒舌囊和第一階段出現。點155的密度在第一階段大幅度的增加。點79在第三階段表現出高量的S35標記。在細線青螺,點40及41的蛋白質被放射線顯影。點262在第二階段出現並持續至第三階段。點53在第三階段表現出高量的S35標記。上述的這些蛋白質可能在齒舌生物礦化的過程中扮演著重要的角色。觀察齒舌發育階段的蛋白質表現的多樣性,有助於進一步探討生物礦化過程中的生物學意義。

The candidate proteins that are involved in biomineralization in the radula of two limpet species C. toreuma and P. striata were studied during different developmental stages by proteome analysis and de novo protein synthesis. Protein patterns revealed by S35-labeling on SDS-PAGE gel differed at different developmental stages in both species, indicating the radulae undergo through some changes and a part of these changes may be related to the biomineralization process. Proteins were also found to be synthesized de novo and transported to the extracellular compartment. Among 1000 and 300 spots resolved in the two-dimensional gels in C. toreuma and P. striata respectively, comparison of different developmental stages of radulae revealed that 19 and 17 protein spots showed differential occurrence at different developmental stages of radulae formation. We found that for example in C. toreuma, a series of spots (spot No.19, 20, 21, 22, 23, 24, 25) and spot 53 appeared only around the radular sac and during stage I. The intensity of spot 155 greatly increased at stage I. Spot 79 presented high intensity 35S-label at stage III. In P. striata, the autoradiography of two protein spots 40 and 41 were found in radular sac. Spot 262 appeared at stage II through early stage III. Spot 53 presented with high intensity 35S-label at stage III. The preceding proteins might play a major role in the mineralization of the radulae. The broad spectrum of protein variability in expression observed during developmental stages of radulae might justify some speculation upon further biological implications of the biomineralization process.

摘要……………………………………………………………..1
Abstract…………………………………………………………2
Introduction……………………………………………………..3
Materials and Methods………………………………………...11
Materials-
Metabolic Labeling with Radionuclides-
SDS-Polyacrylamide Gel Electrophoresis and Autoradiography-
Two-dimensional Electrophoresis and Autoradiography-
Evaluation of 2D data-
Results…………………………………………………………15
SDS-Polyacrylamide Gel Electrophoresis and Autoradiography-
Protein pattern and de novo synthesis differed between stages in C. toreuma-
Protein pattern and de novo synthesis differed between stages in P. striata-
Proteins synthesized de novo and transported to the extracellular compartment-
Two dimensional Electrophoresis and Autoradiography-
Protein spots obtained from the limpet C. Toreuma-
Proteins spots obtained from the limpet P. strata-
Features of 2D- analysis shared by the two limpet C. toreuma and P. striata-
Discussion……………………………………………………24
Chitin and organic matrix framework-
The role of proteins in iron biomineralization-
The role of proteins in silica biomineralization-
Insights for speculation of protein roles in biomineralization-
Acidic macromolecules and biomineralization-
Implications of proteome analysis of SDS-page gel-
Limitations and improvement of experiments-
References……………………………………………………34
Figure Legends……………………………………………….39
Appendix……………………………………………………..92
C. toreuma spot report-
P. striata spot report-

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