[Construction of insecticidal recombinant Bacillus thuringiensis using an integrative vector].
Insecticidal crystal protein gene cry1C which is highly toxic to Spodoptera exigua was cloned into an integrative vector pBMB-F7E, which was derived from Bacillus thuringiensis transposon Tn4430. The recombinant integrative plasmid pBMB-FLC harboring the cry1C gene was gained and transformed into a wild-type Bt strain YBT803-1. A transformant BMB803-A was obtained, and grown at 46 degrees C for about 120 generations, From which three recombinants with cry1C gene integrating into the chromosome were achieved at a frequency of approximately 3.4 x 10(-5). Southern blotting revealed that the integration occurred in different sites of the chromosome. The integrated cry1C were expressed effectively. The results of bioassays showed that the toxicity of recombinants BMB803-X and BMB803-Z against Plutella xylostella were similar to that of strain YBT803-1, and their toxicity to Spodoptera exigua were higher than that of strain YBT803-1.
Yue
Chao-Yin
CY
Huazhong Agricultural University, College of Life Science and Technology, Key Laboratory of Agricultural Microbiology of Ministry of Education, Wuhan 430070, China. yz41@public.wh.hb.cn
Sun
Ming
M
Chen
Shou-Wen
SW
Yu
Zi-Niu
ZN
D000818:Animals; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000254:growth & development; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001427:Bacterial Toxins; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004731:Endotoxins / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D006460:Hemolysin Proteins; D008854:Microscopy, Electron; D010572:Pest Control, Biological / Q000379:methods; D018411:Spodoptera / Q000382:microbiology; D013171:Spores, Bacterial / Q000235:genetics / Q000648:ultrastructure; D014170:Transformation, Genetic
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; C048503:ATGCAT-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific