Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance

被引：150

作者：

Badran, Ahmed H. ^{[1
,2
]}

Guzov, Victor M. ^{[3
]}

Huai, Qing ^{[3
]}

Kemp, Melissa M. ^{[3
]}

Vishwanath, Prashanth ^{[3
]}

Kain, Wendy ^{[4
]}

Nance, Autumn M. ^{[5
]}

Evdokimov, Artem ^{[5
,6
]}

Moshiri, Farhad ^{[5
]}

Turner, Keith H. ^{[5
]}

Wang, Ping ^{[4
]}

Malvar, Thomas ^{[5
]}

Liu, David R. ^{[1
,2
]}

机构：

[1] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02138 USA

[2] Harvard Univ, Howard Hughes Med Inst, Cambridge, MA 02138 USA

[3] Monsanto Co, 245 First St,Suite 200, Cambridge, MA 02142 USA

[4] Cornell Univ, Dept Entomol, Geneva, NY 14456 USA

[5] Monsanto Co, 700 Chesterfield Pkwy West, Chesterfield, MO 63017 USA

[6] HarkerBIO, 700 Ellicott St, Buffalo, NY 14023 USA

来源：

NATURE | 2016年 / 533卷 / 7601期

基金：

美国食品与农业研究所; 美国国家卫生研究院; 美国国家科学基金会;

关键词：

CONTINUOUS DIRECTED EVOLUTION; PROTEIN EVOLUTION; CRYSTAL PROTEIN; CABBAGE-LOOPER; BT CROPS; IN-VIVO; CADHERIN; BINDING; CRY1AC; POPULATION;

D O I：

10.1038/nature17938

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The Bacillus thuringiensis d-endotoxins (Bt toxins) are widely used insecticidal proteins in engineered crops that provide agricultural, economic, and environmental benefits. The development of insect resistance to Bt toxins endangers their long-term effectiveness. Here we have developed a phage-assisted continuous evolution selection that rapidly evolves high-affinity protein-protein interactions, and applied this system to evolve variants of the Bt toxin Cry1Ac that bind a cadherin-like receptor from the insect pest Trichoplusia ni (TnCAD) that is not natively bound by wild-type Cry1Ac. The resulting evolved Cry1Ac variants bind TnCAD with high affinity (dissociation constant K-d = 11-41 nM), kill TnCAD-expressing insect cells that are not susceptible to wild-type Cry1Ac, and kill Cry1Ac-resistant T. ni insects up to 335-fold more potently than wild-type Cry1Ac. Our findings establish that the evolution of Bt toxins with novel insect cell receptor affinity can overcome insect Bt toxin resistance and confer lethality approaching that of the wild-type Bt toxin against non-resistant insects.

引用

页码：58 / +

页数：19

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