Cloning and expression of the insecticidal crystal protein gene Cry1Ca9 of Bacillus thuringiensis G10-01A from Taiwan granaries.
A new cry gene (cry1Ca9) was cloned and sequenced from a Bacillus thuringiensis isolate native to Taiwan (G10-01A). The cry1C-type gene, designated cry1Ca9, consisted of an open reading frame of 3,567 bp, encoding a protein of 1,189 amino acid residues. The polypeptide has the deduced amino acid sequences predicting molecular masses of 134.7 kDa. The gene sequence was compared against the GenBank nucleotide sequence data base. It was found that the cry1Ca9 gene coded for a 134.7-kDa protoxin which had greater than 99.8% homology with the previously reported cry1Ca1 gene, as only three mismatches were found between the two amino acid sequences. When the Cry1Ca9 toxin was expressed in a crystal-negative strain of B. thuringiensis (cryB-), elliptical crystals were produced. Cell extracts from this recombinant strain appear to have high insecticidal activity against lepidopteran larvae (Plutella xylostella).
Kao
Suey-Sheng
SS
Biopesticide Department, Taiwan Agricultural Chemicals and Toxic Substances Research Institute, Council of Agriculture, Wufeng, Taichung Hsien 41301, Taiwan. sskao@tactri.gov.tw
Hsieh
Feng-Chia
FC
Tzeng
Ching-Chou
CC
Tsai
Yeong-Sheng
YS
D000595:Amino Acid Sequence; D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000378:metabolism; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000633:toxicity; D001427:Bacterial Toxins / Q000235:genetics* / Q000633:toxicity; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D004731:Endotoxins / Q000737:chemistry / Q000235:genetics* / Q000633:toxicity; D015870:Gene Expression; D005798:Genes, Bacterial / Q000235:genetics; D006460:Hemolysin Proteins; D007313:Insecta / Q000187:drug effects / Q000254:growth & development; D007814:Larva / Q000187:drug effects / Q000254:growth & development; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010572:Pest Control, Biological; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000633:toxicity; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; D011994:Recombinant Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis