Identification of the minimal active fragment of the Cry1Ah toxin |
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Authors: | Jing Xue Zishan Zhou Fuping Song Changlong Shu Dafang Huang Jie Zhang |
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Institution: | State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, People's Republic of China. |
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Abstract: | cry1Ah1, a novel holo-type gene cloned from Bacillus thuringiensis strain BT-8, encoded a protein exhibiting strong insecticidal activity against lepidopteran insects. To identify the minimal active fragment of the Cry1Ah toxin, 9 pairs of primers were designed to generate different PCR products. Seven PCR products were amplified by different primers using the cry1Ah1 gene as a template and cloned into a pET-21b vector. These positive clones were separately transformed into Escherichia coli. Insecticidal activity against 2nd-instar larvae of Plutella xylostella was performed using the leaf-dip bioassay: the minimal active fragment of the Cry1Ah toxin was located between amino acid residues 50I and 639E. |
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