Construction of ELISA system for detecting Bt toxin resistant insect in the field
田间抗Bt毒素昆虫ELISA检测系统的构建
基本信息
- 批准号:12558069
- 负责人:
- 金额:$ 5.57万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B)
- 财政年份:2000
- 资助国家:日本
- 起止时间:2000 至 2002
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
We have studied for three years to characterize proteins which were specific to either Cry1Ac susceptible P. xylostella or resistant one. These researches have been also done in relation to elucidate the kill mechanism of such B. thuringiensis insecticidal toxins. We have found that the content of oligosaccharylceramide which had longer sugar chain in midgut epithelial cell membrane in the resistant P. xylostella was less than half compared to that of susceptible one. Whereas, another membrane elements such as cholesterol glycerol etc were almost the same each other in both P. xylostella strains. We found a protein whose molecular size and pI were 40 kDa and 5.79, respectively. This was completely missing in Cry1Ac resistant P. xylostella. After completion of cloning of this protein this will be most suitable candidate to make ELISA system to monitor occurrence of resistant insect in field.During these quests, we found various interesting facts. For example, Cry1Ab could kill most effectively the resistant P. xylostella among Cry1A toxins tested, but binding signal of Cry1Ab with apical membrane proteins was least. Cry1Ab bound with 230, 180, 127 and 105 kDa proteins which came from whole midgut membrane. On the other hand, when proteins came from apical-membrane of midgut tissue were used, binding signals were detected in 230, 110, 105, 60 and 55 kDa proteins. These suggested that 230, 110 and 105 kDa proteins which were detected in apical-membrane tissue must localize in apical membrane. Whereas, The proteins of 180 and 127 kDa which were detected in whole midgut tissue but missing in apical membrane materials seemed to localize in basolateral membrane of midgut tissue. These all results suggested that cadherin like protein with 180 kDa molecular size may be in basolateral membrane and could not be Cry toxin receptor.
我们已经研究了三年,以表征蛋白质,无论是敏感的Cry1Ac小菜蛾或抗性之一。这些研究也是为了阐明这种B的杀伤机制。苏云金杆菌杀虫毒素结果表明,抗性小菜蛾中肠上皮细胞膜上糖链较长的寡糖神经酰胺含量较敏感小菜蛾减少一半。而另一种膜成分如胆固醇、甘油等在两个菌株中基本相同。我们发现了一种蛋白,其分子大小和pI分别为40 kDa和5.79。这在抗Cry1Ac的小菜蛾中完全缺失。该蛋白的克隆完成后,将是建立ELISA系统监测田间抗性昆虫发生的最佳候选蛋白。例如,Cry1A毒素对抗性小菜蛾的杀伤效果最好,但与顶端膜蛋白的结合信号最弱。Cry1Ab与来自中肠全膜的230、180、127和105 kDa蛋白结合。另一方面,当使用来自中肠组织顶膜的蛋白时,在230、110、105、60和55 kDa蛋白中检测到结合信号。表明在顶膜组织中检测到的230、110和105 kDa蛋白质必定定位于顶膜。180和127 kDa的蛋白质在整个中肠组织中检测到,而在顶膜材料中缺失,它们似乎定位于中肠组织的基底外侧膜。这些结果提示,分子大小为180 kDa的钙粘蛋白样蛋白可能存在于基底外侧膜,而不是Cry毒素受体。
项目成果
期刊论文数量(38)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
H. Hori, et al.: "Detection of the protoxin of Bacillus thuringiensis serovar japonensis strain Buibui with enzyme-linked immunosorbent assay and its application to the determination of the protein in soil"Appl. Entomol Zool.. 35. 401-411 (2000)
H. Hori 等人:“酶联免疫吸附法检测苏云金芽孢杆菌日本血清型 Buibui 菌株原毒素及其在土壤中蛋白质测定中的应用”
- DOI:
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- 影响因子:0
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N. S. Kumaraswami, et al.: "Analysis and comparison of lipids of brush border membrane vesicle (BBMV) from Plutella xylostella - resistant and susceptible to Cry1Ac δ-endotoxin of Bacillus thuringiensis"Comp. Biochem. Physiol.. 129. 173-183 (2001)
N. S. Kumaraswami 等人:“对苏云金芽孢杆菌 Cry1Ac δ-内毒素具有抗性和敏感性的小菜蛾刷状缘膜囊泡 (BBMV) 的脂质分析和比较”Comp. 129. 173-183 ( 2001)
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- 影响因子:0
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K. Ohtsu, et al.: "Characterization of midgut fluids from Plutella xylostella resistant and susceptible to Cry1Ac toxin of Bacillus thuringiensis"Bulletin of the Faculty of Agriculture, Niigata University. 52. 115-128 (2000)
K. Ohtsu 等人:“对苏云金芽孢杆菌 Cry1Ac 毒素具有抗性和易感性的小菜蛾中肠液的特性”新泻大学农学部通报。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
N.S.Kumaraswami, et al.: "Analysis and comparison of lipids of brush border membrane vesicle (BBMV) from Plutella xylostella -resistant and susceptible to Cry1Acδ-endotoxin of Bacillus thuringiensis"Comp. Biochem. Physiol.. 129. 173-183 (2001)
N.S. Kumaraswami 等人:“对苏云金芽孢杆菌 Cry1Acδ-内毒素具有抗性和敏感性的小菜蛾刷状缘膜囊泡 (BBMV) 的脂质分析和比较”Comp. 129. 173-183 (2001) )
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- 影响因子:0
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- 通讯作者:
Kumaraswami N.S.: "Lipids of brush border membrane vesicles (BBMV) from-Plutella xylostella resistant and susceptible to Cry1Ac O-endotoxin of Bacillus thuringiensis"Comparative Biochemistry and Phyrsiology Part B. (in press). (2001)
Kumaraswami N.S.:“来自小菜蛾的刷状缘膜囊泡 (BBMV) 的脂质对苏云金芽孢杆菌的 Cry1Ac O-内毒素具有抵抗性和敏感性”比较生物化学和生理学 B 部分(正在出版)。
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HORI Hidetaka的其他文献
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{{ truncateString('HORI Hidetaka', 18)}}的其他基金
Recognition mechanism between insect midgut epithelial cell membrane and insecticidal cry toxins
昆虫中肠上皮细胞膜与杀虫叫声毒素的识别机制
- 批准号:
13306006 - 财政年份:2001
- 资助金额:
$ 5.57万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
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小菜蛾(Plutella xylostella)的保幼激素干扰效应
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