Cloning and characterization of two novel crystal protein genes from a Bacillus thuringiensis serovar dakota strain.
Two genes encoding the 32- and 40-kDa polypeptides of Bacillus thuringiensis strain 90-F-45-14 crystals were cloned, expressed in an acrystalliferous B. thuringiensis strain, and sequenced. The polypeptides had deduced molecular weights of 30,319 and 33,885, respectively. The amino acid sequence of the 32-kDa protein was 37.7% identical to the known sequence of a non-insecticidal parasporal protein in B. thuringiensis serovar thompsoni crystals. The sequence of the cloned 40-kDa protein was 37.0% and 22.3% identical to that of the existing Cry protein classes, Cry15Aa1 and Cry23Aa1, respectively. Thus, this protein constitutes a novel protein class, Cry33Aa1. The open reading frames of the two genes were located on the predominant plasmid of 17,629 bp (=11,752 MDa) in the same orientation, and they were separated by the sequence of 32 nucleotides. The two proteins are likely produced simultaneously from a single transcript to form spherical crystals.
Kim
Ho San
HS
Biotechnology & Food Research Institute, Fukuoka Industrial Technology Center, Aikawa-machi 1465-5 Kurume, Fukuoka 839-0861, Japan.
Saitoh
Hiroyuki
H
Yamashita
Satoko
S
Akao
Tetsuyuki
T
Park
Yu Shin
YS
Maeda
Minoru
M
Tanaka
Rie
R
Mizuki
Eiichi
E
Ohba
Michio
M
D000595:Amino Acid Sequence; D001413:Bacillus thuringiensis / Q000166:cytology / Q000235:genetics; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D001427:Bacterial Toxins; D001483:Base Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006460:Hemolysin Proteins; D008957:Models, Genetic; D013171:Spores, Bacterial / Q000648:ultrastructure
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis