博碩士論文 87622011 詳細資訊



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姓名 葉佐平(Zuo-Ping Ye)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 Galectin-1蛋白對昆蟲生長的影響
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摘要(中) GAL1 是一種半乳糖結合蛋白(Galectin)， 具有多重功
能， 包括細胞貼附、增生、分化之功能。GAL1 蛋白於細
胞質中合成後，分泌至細胞外，再與細胞膜上含β-galactosyl
之蛋白結合，而引發細胞內之訊號傳遞。GAL1 與膜蛋白結合的
motif 主要具有 Galβ1-4GlcNAc 結構，與幾丁質聚醣膜(β-1, 4
N-acetyl-D-glucosamine)結構類似；實驗室先前實驗已證實GAL1
可與幾丁質聚醣膜結合。幾丁質為甲殼類生物外骨骼及腸
道外基質(matrix)主要成份；本研究擬探討GAL1 蛋白與幾
丁質之作用基制， 及GAL1 蛋白對昆蟲脫皮之影響； 結果
顯示GAL1 蛋白與幾丁質聚醣膜之作用呈劑量相關， 以
GAL1 蛋白萃取液或純化之GAL1(200-1000µg/ml)餵食昆
蟲幼蟲，可以延遲果蠅幼蟲之發育時間；而小菜蛾幼蟲餵
食GAL1 蛋白後， 幾乎完全無法正常化蛹， 可能幼蟲脫皮
過程受到干擾所致， 然須進一步探討才能確認其作用機
制； 由本實驗之結果顯示GAL1 蛋白可以抑制昆蟲幼蟲正
常發育，具有發展作為生物殺蟲劑之潛力，值得深入探討
GAL1 蛋白作為生物農藥之可行性。
摘要(英) Galectin-1 (GAL1), a β-galactosyl binding lectins, has been shown to
modulate cell adhesion, cell-matrix interaction, cell proliferation and
some immune functions. GAL1 is an extracellular proteins act to
cross-linking cell surface and substrate glycoconjugates. It is proposed
that GAL1, having a single CRD domain recognized the structural motif
Galβ1-4GlcNAc that is similar to that of chitosan membrane (β-1, 4
N-acetyl-D-glucosamine). In an earlier work, we have found that
GAL1-coated chitosan membrane could support 3T3 cells proliferation.
These studies further investigate 1) the interaction of GAL1 and chitosan
in vitro; 2) the effect of GAL1 on insect larvae development. These
results showed that GAL1 dose-dependently bind with chitosan in vitro.
These results suggested that chitosan could be a nature ligand of GAL1.
Furthermore, GAL1 was found to be toxic to some insects, such as the
dipteran of Drosophila melanogaster and the lepidoptera of Plutella
xylostella. The development of diamondback moth (P. xylostella) larvae
were significantly disturbed when feeded with GAL1 extract and/or
recombinant GAL1 proteins purified from GAL1 over-expressed E. coli.
The development of D. melanogaster larvae were also significantly
delayed when feeded with 200-1000µg/ml GAL1; These results
indicate the recombiant GAL1 exhibites high insecticidal
activity and may be a candidate for bio-insecticide.
However, the mechanism of insecticidal toxicity of GAL1
still has to be elucidated
關鍵字(中) ★ 殺蟲劑
★ 昆蟲		 關鍵字(英) ★ Lectin
★ Galectin-1
論文目次 目錄
摘要 I
ABSTRATE II
目錄 III
圖目錄 VI
第一章 緒論 1
1. Lectin 的簡介 1
2. Galectin 的簡介 2
3. 幾丁質 ( Chitin ) 和幾丁聚糖 ( Chitosan ) 的簡介 6
4. 實驗室的初步結果 9
5. 研究動機與目的 11
第二章 材料與方法 13
1. 實驗材料 13
1. 1 菌株 13
1. 2 菌株的培養 13
1. 3 菌株的保存 13
1. 4 抗體 13
1. 5 幾丁聚醣 14
IV
2. 實驗方法 14
2. 1 蛋白質之純化製備 14
2.1.1 蛋白質定量 14
2.1.2 SDS-PAGE 分析（Sodiumdodecyl sulfate polyacrylamide
gel electrophoresis） 14
2. 2 GAL1蛋白質的純化 15
2.2.1 純化GAL1 15
2.2.2 蛋白質的濃縮及冷凍乾燥 16
2. 3 膠體電泳位移分析法（Mobility Shift Assay gel
Eletrophoresis) 16
2. 4 不同濃度GAL1 與幾丁質醣結合測試 17
2. 5 不同濃度的幾丁質醣與GAL 1 結合測試 17
2. 6 乳糖抑制GAL1 與幾丁質醣結合測試 18
2. 7 GAL1 對果蠅生長之測試 18
2. 8 GAL1 對小菜蛾生長之測試 19
第三章、結果 20
1. GAL1 之大量表現與純化 20
2. GAL1 與幾丁聚醣結合移動測試（Mobility shift assay gel
eletrophoresis） 20
V
3. ELISA 測試GAL1 與幾丁聚醣的結合作用 21
4. GAL1 對果蠅的生長測試 22
5. GAL1 對小菜蛾的生長測試 23
第四章、討論 25
1. GAL1與幾丁聚醣之結合作用 26
2. GAL1對昆蟲生長的影響 27
3. 以GAL1 發展為生物農藥 30
第五章、結論及展望 33
第六章、參考文獻 34
附錄 54
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指導教授 黃容南(Rong-Nan Huang) 審核日期 2004-7-19
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