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Construction and characterization of the interdomain chimeras using Cry11Aa and Cry11Ba from Bacillus thuringiensis and identification of a possible novel toxic chimera |
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| Authors: | Yunjun Sun Qiang Zhao Dasheng Zheng Xuezhi Ding Jingfang Wang Quanfang Hu Zhiming Yuan Hyun-Woo Park Liqiu Xia |
| Affiliation: | 1. College of Life Science, Hunan Normal University, Changsha, People’s Republic of China 2. Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, People’s Republic of China 3. Department of Entomology, University of California, Riverside, Riverside, CA, USA 4. Department of Natural and Mathematical Sciences, California Baptist University, Riverside, CA, USA |
| Abstract: | Three structural domains of mosquitocidal Cry11Aa and Cry11Ba from Bacillus thuringiensis were exchanged to produce interdomain chimeras [BAA (11Ba/11Aa/11Aa), ABA (11Aa/11Ba/11Aa), AAB (11Aa/11Aa/11Ba), ABB (11Aa/11Ba/11Ba), BAB (11Ba/11Aa/11Ba), BBA (11Ba/11Ba/11Aa]. Chimeras BAB, BAA, BBA, and AAB formed inclusion bodies in the crystal-negative B. thuringiensis host and produced expected protein bands on SDS-PAGE gel. However, no inclusion body or target protein could be found for chimeras ABA and ABB. In bioassays using the fourth-instar larvae of Culex quinquefasciatus and Aedes aegypti, AAB had ~50 % lethal concentrations of 4.8 and 2.2 μg ml?1, respectively; however, the rest of chimeras were not toxic. This study thus helps to understand the domain-function relationships of the Cry11Aa and Cry11Ba toxins. The toxic chimera, AAB, might be a candidate for mosquito control as its amino acid sequence is different from the two parental toxins. |
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