Directed evolution of the Bacillus thuringiensis insecticidal toxin for the creation of proteinaceous insecticide.
Directed evolution of the Bacillus thuringiensis insecticidal toxin for the creation of proteinaceous insecticide.
批准号:
15380228
负责人:
SATO Ryoichi
金额:
$8.96万
依托单位国家:
日本
项目类别:
Grant-in-Aid for Scientific Research (B)
财政年份:
2003
资助国家:
日本
项目状态:
已结题
起止时间:
2003 至 2004
中文摘要
点击翻译按钮获取中文摘要
英文摘要
In AB-type toxins, which consist of active and binding domains, activity should theoretically be improved if binding affinity to the receptor is improved. The insecticidal toxins (Cry toxins) from B.thuringiensis are AB-type toxins that have been used as organic insecticides and as sources for the development of transgenic plants. In the present report, we describe a method for directed evolution of Cry toxin. This method can be used to increase the binding affinity of Cry toxins for their receptors in target insect pests. Using a commercial T7 phage-display system, we expressed Cry1Aa toxin on the phage surface as fusions with the capsid protein 10B. These recombinant phages bound to a cadherin-like protein that is one of the Cry1Aa toxin receptors in the model target insect Bombyx mori. The affinity of Cry1Aa-expressing phage for the receptor was higher than that of Cry1Ab-expressing phage. Toxin-expressing phages were effectively isolated from T7 wild-type phages using magnetic beads coated with cadherin-like protein. Phages expressing Cry1Aa were isolated from a mixed suspension of phages expressing Cry1Ab and concentrated by up to 1.3 x 10^<5->fold. Finally, random mutations were made in amino acid residues 369-375 in domain 2 of Cry1Aa toxin, the mutant toxins were expressed on phages, and the resulting phage library was screened five times with cadherin-like protein-coated beads. As a result, phages expressing abnormal or low-affinity mutant toxins were excluded, and phages with high-affinity mutant toxins were selected. The insecticidal activity of one of these toxins against B.mori was increased four-fold over that of wild-type Cry1Aa toxin. These results indicate that a method combining T7 phage display with selection using cadherin-like protein-coated magnetic beads can be used to increase the activity of easily obtained, low-activity Cry toxins from bacteria.
期刊论文(5)
专著(0)
科研奖励(0)
会议论文
目的昆虫に効くδ内毒素変異体を選抜するためのシステム
用于选择对目标昆虫有效的δ内毒素突变体的系统
DOI:
--
发表时间:
2005
期刊:
影响因子:
--
作者:
[]
通讯作者:
Location of the Bombyx mori aminopeptidase N type l binding site on a Bacillus thuringiensis CrylAa toxin.
苏云金芽孢杆菌 CrylAa 毒素上的家蚕氨肽酶 N I 型结合位点的位置。
DOI:
--
发表时间:
2005
期刊:
Applied and Environmental Microbiology 71
影响因子:
--
作者:
[S.Atsumi他 8名]
通讯作者:
S.Atsumi他 8名
Location of the Bombyx mori Aminopeptidase N type 1 binding site on a Bacillus thuringiensis Cry1Aa toxin.
苏云金芽孢杆菌 Cry1Aa 毒素上家蚕氨基肽酶 N 1 型结合位点的位置。
DOI:
--
发表时间:
2005
期刊:
Appl Environ Microbiol. 71(7)
影响因子:
--
作者:
[Shogo Atsumi, Eri Mizuno, Hirotaka Hara, Kazuko Nakanishi, Madoka Kitami, Nami Miura, Hiroko Tabunoki, Ayako Watanabe, Ryoichi Sato]
通讯作者:
Ryoichi Sato
Evolvement of "durability mechanics" for the evaluation of shrinkage effect on performance of reinforced concrete beams
-
批准号:24246077
-
项目类别:Grant-in-Aid for Scientific Research (A)
-
资助金额:$21.8万
-
财政年份:2012
-
负责人:SATO Ryoichi
-
依托单位:
DEVELOPMENT OF A PRACTICAL TENSILE CREEP MODEL BASED ON SIMPLE EXPERIMENTS
-
批准号:24656272
-
项目类别:Grant-in-Aid for Challenging Exploratory Research
-
资助金额:$1.5万
-
财政年份:2012
-
负责人:SATO Ryoichi
-
依托单位:
Generation of a new category, "Proteinaceous Insecticide" based on mechanisms which cause specificity diversity of Bacillus thuringiensis insecticidal protein.
-
批准号:24310054
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$11.32万
-
财政年份:2012
-
负责人:SATO Ryoichi
-
依托单位:
Development of Accurate Detection/Identification Method of Stricken Residential Buildings Using Radar Porlarimetry
-
批准号:22510188
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$2.91万
-
财政年份:2010
-
负责人:SATO Ryoichi
-
依托单位:
Realization of a secure and safe proteinaceous-insecticide based on the directed evolution of the insecticidal toxin from soil bacterium, Bacillus thuringiensis
-
批准号:21310051
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$11.81万
-
财政年份:2009
-
负责人:SATO Ryoichi
-
依托单位:
Durability Mechanics for High Shrinkage Concrete Members
-
批准号:21246071
-
项目类别:Grant-in-Aid for Scientific Research (A)
-
资助金额:$25.46万
-
财政年份:2009
-
负责人:SATO Ryoichi
-
依托单位:
Detection and Classification of Landslide and/or Stricken Residential Area in Mountainous Region Using Radar Polarimetry
-
批准号:19510183
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$2.91万
-
财政年份:2007
-
负责人:SATO Ryoichi
-
依托单位:
Development of Expansive-High Strength Concrete and Systematic Evaluation of Structural Behavior of Reinforced Concrete Beams
-
批准号:16360220
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$8.83万
-
财政年份:2004
-
负责人:SATO Ryoichi
-
依托单位:
Approach for creation of proteinaceous insecticide from Bacilhus thuringiensis delta-endotoxin by directed evolution using phage display system
-
批准号:12558068
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$8.45万
-
财政年份:2000
-
负责人:SATO Ryoichi
-
依托单位:
Research for a basic anti-microbial recognition system of the animal using silkworm, Bombyx mori
-
批准号:10680601
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$1.98万
-
财政年份:1998
-
负责人:SATO Ryoichi
-
依托单位:
Creation of proteinaceous insecticide from Bacuillus thuringlensis δ-endotoxin and single chain variable fragment
-
批准号:09558076
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$5.7万
-
财政年份:1997
-
负责人:SATO Ryoichi
-
依托单位:
Time-Dependent Deformations of Prestressed and Prestressed Reinforced High Strength Concrete Members
-
批准号:08455204
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$2.24万
-
财政年份:1996
-
负责人:SATO Ryoichi
-
依托单位:
Characterization of recognition specificity and analysis of variety of recognition proteins of BT toxin.
-
批准号:08660408
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$1.54万
-
财政年份:1996
-
负责人:SATO Ryoichi
-
依托单位:
Study on the old : strata of the accents around Tokyo
-
批准号:07610422
-
项目类别:Grant-in-Aid for Scientific Research (C)
-
资助金额:$1.22万
-
财政年份:1995
-
负责人:SATO Ryoichi
-
依托单位:
Engeineering of delta-endotoxin from Bacillus thuringiensis to make a new specificity.
-
批准号:05660385
-
项目类别:Grant-in-Aid for General Scientific Research (C)
-
资助金额:$1.34万
-
财政年份:1993
-
负责人:SATO Ryoichi
-
依托单位:
DESIGN METHOD OF REINFORCED CONCRETE MEMBERS WITH THERMAL CRACKING
-
批准号:03555105
-
项目类别:Grant-in-Aid for Developmental Scientific Research (B)
-
资助金额:$1.92万
-
财政年份:1991
-
负责人:SATO Ryoichi
-
依托单位:
STUDIES ON THE EFFECTS OF ENVIRONMENTAL CONDITION ON LONG-TERM DEFORMATION AND CRACK WIDTH OF REINFORCED CONCRETE MEMBERS
-
批准号:03650389
-
项目类别:Grant-in-Aid for General Scientific Research (C)
-
资助金额:$1.28万
-
财政年份:1991
-
负责人:SATO Ryoichi
-
依托单位:
A Study of Properties and Prediction of Long-term Deformation of Reinforced Concrete Members
-
批准号:01550372
-
项目类别:Grant-in-Aid for General Scientific Research (C)
-
资助金额:$1.09万
-
财政年份:1989
-
负责人:SATO Ryoichi
-
依托单位:
海外基金