• Journal of Microbiology and Biotechnology
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• v.5 no.3
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• pp.138-142
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• 1995

Bacillus thuringiensis strain BT-209 was isolated from a soybean grain dust sample in Korea. The strain BT-209 produced two different sizes of cuboidal crystals and one spore in the cell. In the biochemical characterization, the strain BT-209 showed negative reactions on the production of urease, and the utilization of citrate and sucrose. Examination of its antibiotic resistance revealed that while the strain BT-209 showed higher sensitivity than B. thuringiensis subsp. kurstaki HD-1 to ampicillin, bacitracin, chlortetracycline, gentamycin, neomycin, penicillin G, tetracycline and tobramycin, it was more resistant to methicillin than B. thuringiensis subsp. kurstaki HD-1. The $\delta$-endotoxin crystal of strain BT-209 consisted of three proteins with apparent molecular weights of appoximately 148, 135 and 62 kDa on a 10% SDS-PAGE. The strain BT-209 had at least eight different plasmids with sizes of 4.1, 5.2, 6.3, 8.6, 14.6, 24.5, 67.6 and 77.6 Kb. The strain BT-209 showed strong lethalities of 70% and 87% against Bombyx mori and Hyphantria cunea larvae. at 72 h, respectively.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.2
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• pp.183-186
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• 2004

Recently, we have developed an easy, simple and convenient circular DNA cloning system named plasmid capture system (PCS). To investigate usefulness of PCS in cloning of plasmids from Bacillus thuringiensis strains, PCS donors, pPCS-S and pPCS-L were applied to clone plasmids of B. thuringiensis subsp. israelensis by in vitro transposition using 4{TnsABC^*}$ transposase. In result, 3 small plasmids were cloned, and these were consistent with pTX14-1, pTX14-2 and pTX14-3 reported previously from B. thuringiensis subsp. israelensis. Therefore, the PCS can be successfully applied to clone small plasmids from B. thuringiensis strains.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.185-189
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• 2001

Wild type Bacilus thuringiensis subsp. israelensis and B. sphaericus toxins have been used separately as active in ingredients for bacterial insecticides to control mosquito larvae due to their comparable toxicity to chemical insecticides. Cry11B, recently cloned from B. thuringiensis subsp. jegathesan, shows higher toxicity against three major species of mosquito larvae than Cry11A, one of the major component of B. thuringiensis subsp. israelensis inclusion body. To determine whether the combination of cry11B and B. sphaericus binary toxins is as toxic as B. thuringiensis subsp. israelensis parental strain, cry11B and B. sphaericus binary toxins genes were co-expressed as an operon using cytlA promoters/STAB-SD hybrid expression system in B. thuringiensis subsp. israelensis acrystalliferous strain 4Q7. However, unexpectedly, B. sphaericus binary toxins were barely produced, whereas relatively large amount of Cry11B was produced. When this strain was grown in four different media, NB+G and Peptonized Milk produced more toxin proteins and spores per unit of media than GYS and G-Tris. Toxicity of this strain against fourth instar Culex quinquefasciatus was ranged from of 8.3 to 45.7 ng/ml, with NB+G culture being the highest, and GYS culture was the lowest.

• Korean journal of applied entomology
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• v.32 no.4
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• pp.426-432
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• 1993

Thirteen isolates of Bacillus thunngiensls producing parasporal mclusions, obtained from 45 samples of dust and soil of sericultural tarms in Kyeong-ki province, were exammed for their toxicity against larvae of Lepidoptera, Dipwra and Coleoptera. Of these isolates, Bacillus f thuringiensis NT0423 was toxic to bath Lepidoptera, and Dipteran larvae. NT0423 showed that the $LC_{50}$ values against the Lepldaptora, Plutella macuhpennis and the Diptera, Culex pipiens were $1.30\times10^6$ CFU/ml, $2.88\times10^5$ CFU/ml, respectively. The tYPlcal bipyramldal crystals produced by NT0423 composed of protoxms of 130-140kDa encoded by one or more plasm mid-horne genes. Also, plasmid DNA analysis indicated Lhat this isolate has 9 plasmids which d differ with reported several B. thuringiensis strains.

• Journal of Microbiology and Biotechnology
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• v.4 no.4
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• pp.322-326
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• 1994

Bacillus thuringiensis strain BT-14 was isolated from alfalfa dust in Korea. The strain BT-14 produced one bipyramidal crystal and one spore in the cell. The biochemical characteristics of the strain BT-14 were similar to those of Bacillus thuringiensis subsp. kurstaki HD-l. Examination of its antibiotic resistance revealed that while the strain BT-14 was less resistant than BTK HD-l to ampicillin, gentamycin, neomycin and tobramycin, it was more resistant to amikacin than BTK HD-l. The $\delta$-endotoxin crystal of strain BT-14 consisted of a single protein with a high molecular weight of ca 135 KD on a 10% SDS-PACE. The strain BT-14 contained at least nine different plasmids with sizes of 2.9, 5.3, 5.8, 6.2, 9.4, 15.1, 18.1, 23.1 and 79 Kb. In insect bioassay, the isolated strain BT-14 showed lethality of 67% against Plutella xylostella larvae at dilution of 5$\times$$l0^{-4}$ (5$\times$l0 to 3$\times$$l0^2$ spores/ml), which is, almost equivalent to that of BTK HD-l.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.361-367
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• 2010

Bacterial contamination of cooked rice was analyzed to evaluate the microbial safety. Thirty raw rice samples were collected in Korea and cooked in an electric rice cooker. Mesophilic aerobe, food-poisoning Bacillus cereus group, and their toxin genes were determined on cooked rice. The percentage of total mesophilic aerobe based on 1-3 log CFU/g was 27% among the samples. Bacillus spp. in MYP selective medium was similar to the number of mesophilic aerobe, whileas Bacillus spp. was detected in most samples after enrichment. Thirty-seven isolates from 30 cooked rices were identified as B. thuringiensis, B. cereus, B. valismortis, B. pumilus, B. coagulans, B. licheniformis, Geobacillus stearothermophilus, and Brevibacillus laterosporus. Twenty isolates (54%), more than half of the isolates, were B. thuringiensis while nine (27%) were identified as B. cereus. All B. thuringiensis isolates possessed non-hemolytic toxin genes and interestingly, seven B. cereus among nine isolates possessed emetic toxin genes. More B. thuringiensis was present on the cooked rice than B. cereus and most B. cereus possessed emetic toxin genes rather than diarrheal toxin genes. Therefore, food-borne outbreak due to B.cereus on the cooked rice kept at room temperature might be examples of emetic food-poisoning.

• Korean journal of applied entomology
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• v.25 no.2 s.67
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• pp.71-76
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• 1986

Bacillus thuringiensis var. thuringiensis H1 (BTT) strain was cultured in the 4 different fermentation media and then measured their growths and the productions of endotoxin crystals from the culture media. Out of the 4 media, the productions of the endotoxin crystals and spores were maximal in the pH9-M-3 medium. The wet weight of BTT cells grown in the 150ml culture was approximately 3.218g and the number of the viable spores was $3.3{\times}10^{10}/ml$, and the ratio of the endotoxin weight over total cell weight was 20.05%. The generation time of the BTT bacteria in the M-1 was about 47.6 minutes in the M-2, 132.9 minutes in the M-3 and 110.2 minutes in the M-4. The proper pH for the production of the endotoxin by BTT appeared to be pH 6.5.

• The Korean Journal of Pesticide Science
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• v.18 no.3
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• pp.181-190
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• 2014

An entomopathogenic bacterium, Bacillus thuringiensis (Bt), can sporulate along with production of insecticidal Cry toxins. Bt Cry toxins exhibit relatively narrow spectrum to target insects due to their specific interactions with midgut receptors. This study designed several strategies to enhance Bt efficacy in target insect spectrum and insecticidal activity. Four Cry toxins were purified from four different Bt strains and showed relatively narrow target insect spectrum. However, the Cry mixtures significantly expanded their target insect spectra. The additional effect of baculovirus to Cry toxin was tested with recombinant baculoviruses expressing Cry1Ac or Cry1Ca. However, the baculovirus was little effective to expand target insect spectrum. Bacterial culture broth of Xenorhabdus nematophila (Xn) significantly suppressed insect cellular immune response and increased Cry toxicity. The addition of Xn culture broth to Cry mixture significantly enhanced Bt efficacy in target insect spectrum and insecticidal activity.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.2
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• pp.123-129
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• 2000

Expression of the crylAcl gene of an acrystalliferous Bacillus thuringiensis strain under the control of the native or $\alpha$-amylase gene promoter was investigated. The crylAcl gene was cloned in a B. thuringiensis - E. coli shutle vector, pHT3101, undder the control of either the native promoter (pProAc) or the $\alpha$-amylase promoter from Bacillus subtilis (pAmyAc). These two recombinant plasmids were successfully expressed in B. thuringiensis subsp. kurstaki Cry B. The first transformant (ProAc/CB), harboring pProAc, expressed an about 130 kDa protein begining 24 hr after inoculations just as in the case of the wild type of B. thuringiensis subsp. kurstaki HD-73. The second pAmyAc-transformant (AmyAc/CB) began to express the gene just 6 hr after inoculation, but Western analysis showed that the activity of the $\alpha$-amylase promoter was relatively weaker than that of the native promoter. As expected, their toxicity against Plutella xylostella larvae was dependent on the amount of Cry1Acl protein expressed.

• The Korean Journal of Pesticide Science
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• v.14 no.4
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• pp.446-455
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• 2010

The characteristics of parasporal inclusion body from Bacillus thuringiensis subsp. kurstaki KB099 isolate which is high bioactive to the tobacco cutworm, Spodoptera litura, were examined. Parasporal inclusion of B. thuringiensis subsp. kurstaki KB099 isolate showed only 1 band at 130 kDa compared with B. thuringiensis subsp. kurstaki HD-l isolate producing 2 protein bands at 130 kDa and 60 kDa from by SDS-PAGE analysis without any enzyme treatment. Also, we confirmed that gut extract of sensitive S. litura KB099 isolate had digested only 60 kDa ${\delta}$-endotoxin protein. When the digestive enzyme of sensitive insect responsible for parasporal inclusion from KB099 and HD-l isolate was treated to each of them, protein band 60 KDa of KB099 was maintained up to 12 hours but all bands of HD-l were disappeared within 6 hours. In KB099 isolate, 6 genes (Cry1Aa, Cry1Ab, Cry1Ac, Cry1C, Cry1D and Cry1I) were identified by PCR analysis. Also, $Cry^-$ mutant of KB099 isolate was investigated by phase- contrast microscope, SDS-PAGE and PCR.