• 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.

• Korean journal of applied entomology
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• v.33 no.3
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• pp.178-183
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• 1994

The prolein components of each crystal toxin of Bacillus thuringiensis kurstaki and alzawai were separated by SDS-PAGE. The major clqistal proteins from both shins were composed of paiypeptides having molecular weights of 130 kd and 64 kd. Digestive mixture of both. toxins with t~ypsin and gut juices shared 62 kd polypeptide which may be major actwe toxh. However, aizawal produced much less amount of 62 kd polypeptide than kurstaki did. On ingestion with Bt &-endotoxin, larvae of Hyphantria cunea developed 45 kd stress protein in the several tissues including fat body, which was induced by heat and cold shock.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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• pp.315-319
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• 1985

Delta-endotoxin crystals of B. thuringiensis var. kurstari and B. thuringiensis var. israelensis were purified by NaBr density gradient centrifigation and the wet weight of the BTK endotoxin was approximately 23.79％ of the cell wet weight and that of BTI was 25％. The shape of BTK crystal was bipyramidal, whose size was 1.7${\mu}{\textrm}{m}$ $\times$ 0.9${\mu}{\textrm}{m}$ and that of BTI was a spheroid, whose size was about 1.6$\times$0.45${\mu}{\textrm}{m}$. The molecular weight of BTK crystal protein was approximately 134,000 daltons and that of BTI was about 128,000 daltons.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.168-173
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• 1988

Immunological analysis between endotoxin proteins produced by Bacillus thuringiensis serovar. kurstaki HD1 and HD73 have been investigated by using polyclonal antibodies. The antisera against the endotoxin proteins were prepared from rabbits injected with the endotoxin protein antigens. When about 2mg/$m\ell$ of the antigens were injected for 7 times, the titers were highest. The stability of the antigens was reduced to about 50% after 9 days incubation at 4$^{\circ}C$. The sensitibity of endotoxin protein from B. thuringiensis HD1 and HD73 by indirect ELISA was 50ng/$m\ell$ and 400ng/$m\ell$, respectively. The cross reaction of antiserum appeared that anti-HD1 partialy reacted with crystal protein but anti-HD73 reacted with HD1 endotoxin about 100%.

• 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.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.658-663
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• 2007

Bacillus thuringiensis is a well-known species of entomophathogenic bacteria that is widely used as a biopesticide against many insect pests. It produces parasporal crystals ($\delta$-endotoxin) and endospores during sporulation. In this report, the $\delta$-endotoxin produced by Bacillus thuringiensis BT-1 and BT-2 were characterized by Scanning Electron Microscope(SEM), Transmission Electron Microscope(TEM), SDS-PACE, and solubilization activity by alkaline solution. BT-1, BT-2 were cultured in the GBY medium, and the $\delta$-endotoxin of them was purified with discontinuous sucrose density gradient centrifugation. Their $\delta$-endotoxin was observed by SEM and TEM. Morphologically, the $\delta$-endotoxin of BT-1 was a square and flat type, whose size was $1.73{\mu}m{\times}0.7{\mu}m$, and the $\delta$-endotoxin of the BT-2 was spherical form whose size was $1.1{\mu}m{\times}0.9{\mu}m$ determined by SEM and TEM. The $\delta$-endotoxin of the BT-1 was composed of 28 kDa and 21 kDa, however, it of the BT-2 was composed of 50 kDa, 35 kDa, and 22 kDa bands determined by SDS-PACE. The purified crystals of BT-1 and BT-2 were dissolved gradually in alkaline solution as time goes by, and it was perfectly dissolved after 3 hours. It is supposed that the $\delta$-endotoxin of crystal was converted to a state of activation in the course of time in the intestines of insect.

• Microbiology and Biotechnology Letters
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• v.13 no.1
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• pp.51-57
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• 1985

Delta-endotoxin in Bacillus thuringiensis var. finitimus, HD-1, HD-9 and HD-73 strains were isolated by NaBr, CsCl and Renografin density gradients. The purity of the toxin was about 98%. The purified o-endotoxin was analyzed by SDS-PAGE, electron microscopic observation and bioassay. According to SDS-PAGE, the molecular weight of subunits of the o-endotoxin were about 66,000 and 130,000 daltons. The shapes of the crystal toxin observed by TEM except finitimus strain were bipyramidal. When the purified endotoxin was bioassayed against tobacco horn worm, the entomocidal activities ($1{\mu}g/ml$) of HD-1 and HD-73 strains were, respectively, 60% and 100% at nine days after treated. The molecular weights of the plasmids isolated from B. thuringiensis were various from 0.5 to 120 Kb. The numbers of plasmids in HD-1, HD-9 and HD-73 strains were 12, 3 and 11, respectively, but B. thuringiensis var. finitimus strain had no plasmid.

• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.534-540
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• 1999

Thirty-seven strains of Bacillus thuringiensis were isolated from Korean soil and examined for H-antigen serotyping, toxicity, and different spectra of biological activities. The isolate HL-175 bore a specific H-antigen, different from the 51 known serotypes, a spherical $\delta$-endotoxin crystal, and minor different biochemical characteristics. It was resistant to ampicillin, colistin, and penicillin G. Therefore, it was classified as a new serotype, H52, with the name kim. The other 36 isolates also produced endotoxin crystals and endospores. The crystal shape of eight strains was cuboidal while the others were bipyramidal. Biochemical characteristics of the isolates were only slightly different from the known serotypes of B. thuringiensis. The flagellar (H) antigens of the 36 isolates were identified as: one colmeri (H21), three galleriae (H5a,5b); two pakistani (H13); one toumanoffi (H11a, 11b); and twenty-nine kurstaki (H3a,3b). All 36 isolates were resistant to ampicillin, colistin, penicillin, cephalothin, and chloramphenicol.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.647-652
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• 1990

The production of delta-endotoxin crystal and the cloning of endotoxin protein gene in Bscillus thuringiensis subsp. kurstaki HD1 strain were studied. The strain produced bipyramidal crystals ($2.9\times 1.0 \mu m$) in their cells during sporulation. The B. thuringiensis contained about 10 plasmid DNA elements ranging from 2.1 to 80 kilobases. The 73 kb plasmid DNA, the 29 kb BamHI fragment and the 7.9 kb Pstl DNA fragment hybridized to the pHL probe. The 7.9 kb fragment was eluted and cloned in the PstI site of pBR322 vector and transformed into E. coli HB101, which produced insecticidal proteins killing Bornbyx mori larvae.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1106-1110
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• 2001

The parasporal biogenesis of crystal inclusion during the sporulation of Bacillus sphaericus strain 1593 was observed using transmission electron microscopy. The crystal biogenesis and sporulation process involved a sequence of events talking about 10 h. The sporulation Precesses were found to be similar to previous findings. The crystal biogenesis of B. sphaericus was initiated at the start of engulfment and nearly completed by the time of exosporium formation. The crystal formation was clearly associated with the outer forespore membrane from stages III through VI, and the crystals grew from polypeptide-like chains originated from the outer forespore membrane. These observations are different from previous findings, which report no association with the forespore membrane. The crystals were located adjacent to the outer membrane of the spore until the release stage. The axes size of the bipyramidal crystal was approximately $0.25{\mu}m{\times}42{\mu}m$. During crystal biogenesis, the crystal development could be classified into four stages; initiation stage Cl (sporulation stage . III), growth stage C2 (sporulation III to V), envelopment and maturation C3 (sporulation V to V), and finally release stage C4 (sporulation Vll).