[Expression characteristic of Bacillus thuringiensis cry1 gene in Pseudomonas fluorescens Pfx-18].
The plasmids of lepidopteran-specific Bacillus thuringiensis strain from our laboratory were hybridized with RNA probe of cry 1Aa EcoR I-F fragment labelled using DIG. Cry1 gene was located in 39.3 MD plasmid. The plasmid was digested with Hind III and analysed by southern blot. It appeared both 7.1 kb and of 6.5 kb positive bands. The 7.1 kb fragment was ligated to broad-host-range vector pSUP106 and transformed into Pseudomonas fluorescens Pfx-18. The cloned strain, LZP-1 was obtained. The plasmids of LZP-1 were analysed by PCR. The results showed that gene-type is cry1Ab. SDS-PAGE analysis demonstrated that LZP-1 could express 66 kD insecticidal crystal protein and some small molecular weight peptides. Bioassay showed that motality of 1000-fold diluted fermentation broth was 33% to 3rd instar plulella xyloslelly larvae.
Liu
Z
Z
Key Lab. of Agri-Microbiology, Ministry of Agriculture, Dept. of Microbial Science and Technology, Huazhong Agricultural University, Wuhan 430070.
Tang
J
J
Yu
L
L
Sun
M
M
Yu
Z
Z
D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000633:toxicity; D001427:Bacterial Toxins / Q000235:genetics* / Q000633:toxicity; D003001:Cloning, Molecular; D004731:Endotoxins / Q000235:genetics* / Q000633:toxicity; D006460:Hemolysin Proteins; D007814:Larva / Q000187:drug effects; D007915:Lepidoptera / Q000187:drug effects; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011551:Pseudomonas fluorescens / Q000235:genetics*
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis