Analysis of Biological Domains in Insecticidal Protein from B. Thuringiensis

苏云金芽孢杆菌杀虫蛋白的生物结构域分析

基本信息

  • 批准号:
    01560105
  • 负责人:
  • 金额:
    $ 1.34万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
  • 财政年份:
    1989
  • 资助国家:
    日本
  • 起止时间:
    1989 至 1990
  • 项目状态:
    已结题

项目摘要

Bacillus thuringiensis var. Israelensis (Bti) produces insecticidal crystalline protein (ICP), delta-endotoxin, which specifically kills dipteran insect larvae, mosquito and black fly. The ICP have several toxicities, that is, 1) dipteran insecticidal, 2) insect cell toxic, 3) insect specific neurotoxic, 4) mammalian cell toxic, 5) mammalian hemolytic and 6) suckling mouse toxic activities. The ICP senes encode 128 kDa (ISRH3) and 135 kDa (ISRH4) proteins and determined as respects the nucleotide sequences. The ISRH3 gene is consisted of 3405 by encoding 1135 amino acid. Then in this project, we attempt to identify the active domain in ISRH3 protein on the activities.Several mutants of ISRH3 deleted from C of N terminal were constructed by treatment of several restriction enzymes and the proteins from the genes were assayed by the mosquito larvae and an established insect cell (TN-368 ). The protein produced by the gene encoding from N terminal to No. 635 amino acid has also insecticid … More al and insect cytolytic activities. However, the peptide from N terminal to No. 602 loses these two activities. The proteins of No. 613 - No. 1135 amino acid (C terminal) have the two activities, though the proteins of No. 632 or No. 641 - C terminal showed no clear results as to the activities. Then that protein of No. 716 - No. C terminal had lost the activities. The ISRH3 proteins deleted a central parts No. 560 - No. 772 amino acid had also lost its. The several small peptides central parts, No. 560 - No. 820 had lost the activities. From these results the central parts included No. 613 - No. 635 amino acid is one of domain and another domain (R or R') locate at N or C terminal, respectively. It was suggested that the domains for insecticidal and insect cell toxic activities required two domains, that is, A and another R or R' domains and also suggested the insecticidal activities was the insect cell toxic activity. However, it is ISRH3 does not have the mammalian cell toxic, hemolytic activities and the suckling mouse toxicity. The epitop of an antibody against ISRH proteins recognizes amino acid sequences around No. 773 - No. 820. Less
苏云金杆菌Israelensis(Bti)产生杀虫晶体蛋白(ICP),δ-内毒素,其特异性地杀死双翅目昆虫幼虫、蚊子和黑蝇。ICP具有多种毒性,即1)双翅目杀虫活性,2)昆虫细胞毒性,3)昆虫特异性神经毒性,4)哺乳动物细胞毒性,5)哺乳动物溶血性和6)乳鼠毒性。ICP基因编码128 kDa(ISRH 3)和135 kDa(ISRH 4)蛋白质,并根据核苷酸序列确定。ISRH 3基因由3405个氨基酸组成,编码1135个氨基酸。本研究利用多种限制性内切酶对ISRH 3蛋白进行了酶切,构建了ISRH 3蛋白的C/N端缺失突变体,并利用蚊幼虫和已建立的昆虫细胞(TN-368)对突变体的蛋白质进行了分析。编码N端至第635位氨基酸的基因所产生的蛋白质也已被纯化, ...更多信息 al和昆虫细胞溶解活性。而从N端到602号的肽段则失去了这两种活性。第613 - 1135位氨基酸(C端)的蛋白质具有这两种活性,而第632或641 - C端的蛋白质的活性没有明确结果。716 -C端蛋白丧失活性。ISRH 3蛋白缺失了中心部分第560 - 772位氨基酸,也失去了其功能。中心部分的几个小肽,No.560-No.820已失去活性。从这些结果可以看出,中心部分包括第613 - 635位氨基酸,分别是位于N端和C端的一个结构域和另一个结构域(R或R ')。结果表明,杀虫活性和昆虫细胞毒活性的结构域需要两个结构域,即A和另一个R或R'结构域,杀虫活性即昆虫细胞毒活性。然而,ISRH 3不具有哺乳动物细胞毒性、溶血活性和乳鼠毒性。抗ISRH蛋白的抗体的表位识别No.773-No.820附近的氨基酸序列。少

项目成果

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科研奖励数量(0)
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HIMENO Michio其他文献

HIMENO Michio的其他文献

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{{ truncateString('HIMENO Michio', 18)}}的其他基金

ISOLATION AND ANALYSIS OF RECEPTOR FOR INSECTICIDAL PROTEIN, delta-ENDOTOXIN
杀虫蛋白δ-内毒素受体的分离与分析
  • 批准号:
    04660093
  • 财政年份:
    1992
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
A Trial for a Creation of a New Protein Fiber by Modification of Fibroin Gene, Bombyx mori L.
通过修饰丝素基因(Bombyx mori L.)创建新蛋白纤维的试验。
  • 批准号:
    62480046
  • 财政年份:
    1987
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)

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基于苏云金芽孢杆菌杀虫蛋白特异性多样性的机制,产生了一个新类别“蛋白质杀虫剂”。
  • 批准号:
    24310054
  • 财政年份:
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    $ 1.34万
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    Grant-in-Aid for Scientific Research (B)
Nonaqueous chemical modification of lyophilized proteins: application to be insecticidal protein from bacillus thuringiensis
冻干蛋白的非水化学修饰:苏云金芽孢杆菌杀虫蛋白的应用
  • 批准号:
    90550-1998
  • 财政年份:
    2001
  • 资助金额:
    $ 1.34万
  • 项目类别:
    Discovery Grants Program - Individual
Analysis of a novel mode of action of the dipteran-specific insecticidal protein
双翅目特异性杀虫蛋白的新作用模式分析
  • 批准号:
    13836008
  • 财政年份:
    2001
  • 资助金额:
    $ 1.34万
  • 项目类别:
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  • 批准号:
    90550-1998
  • 财政年份:
    2000
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    $ 1.34万
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    Discovery Grants Program - Individual
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冻干蛋白的非水化学修饰:苏云金芽孢杆菌杀虫蛋白的应用
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  • 财政年份:
    1999
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    $ 1.34万
  • 项目类别:
    Discovery Grants Program - Individual
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冻干蛋白的非水化学修饰:苏云金芽孢杆菌杀虫蛋白的应用
  • 批准号:
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    1998
  • 资助金额:
    $ 1.34万
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    Discovery Grants Program - Individual
Studies on Development of Efficient Transgenic System for Alien Gene of Insecticidal Protein in Sugarbeet
甜菜杀虫蛋白外源基因高效转基因体系的开发研究
  • 批准号:
    09460001
  • 财政年份:
    1997
  • 资助金额:
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生产具有杀虫蛋白基因的抗虫转基因草坪植物。
  • 批准号:
    07456002
  • 财政年份:
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ISOLATION AND ANALYSIS OF RECEPTOR FOR INSECTICIDAL PROTEIN, delta-ENDOTOXIN
杀虫蛋白δ-内毒素受体的分离与分析
  • 批准号:
    04660093
  • 财政年份:
    1992
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    $ 1.34万
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LOCALIZATION OF TOXICITY DOMAINS OF B THURINGIENSIS INSECTICIDAL PROTEIN
苏云金芽孢杆菌杀虫蛋白毒性域的定位
  • 批准号:
    3957949
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