Identification of the minimal active fragment of the Cry1Ah toxin.
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.
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.
Jing
J
Xue
Zishan
Z
Zhou
Fuping
F
Song
Changlong
C
Shu
Dafang
D
Huang
Jie
J
Zhang
D000595:Amino Acid Sequence; D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D007915:Lepidoptera / Q000187:drug effects*; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
D004731:Endotoxins; D011994:Recombinant Proteins