• 한국미생물·생명공학회지
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• 제30권3호
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• pp.230-234
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• 2002

모기 살충성 Bacillus thuringiensis subspangiensis 21-2균주의 용혈성 내독소 단백질의 특성을 검토하고자 21-2균주의 용혈성을 가진 유전자를 Escherichia coli HBIO쎄 형질전환시켰다. 이들 중에서 형질전환 균주 47은 독소 단백질을 생산하며 2.5 kb DNA을 함유한다는 것을 효소항체법, immunoblot및 DNA전기영동법으로 확인하였다. 또한, 형질전환 균주 47-5는 2.5 kb DNA를 다시 Hind ll견 절단하여 pUC118 연결시켜서 조제하였다 그 결과형질전환 균주 47-5은 1.Bkb DNA를 함유하며, 23 kD꺼 독소 단백질을 발현하고, 발현된 독소 단백질은 Aedes aegypti모기 유충에 독성을 나타내었다. 또한 23 kDa의 내독소 단백질 그 자체로는 사람의 적혈구를 용해하지 못하였으나, proteinase K로 처리한 후에는 적혈구에 대해 용혈성을 나타내었다.

• 미생물학회지
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• 제53권3호
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• pp.216-218
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• 2017

생물학적방제 효과가 뛰어난 Bacillus thuringiensis C25 균주의 유전체 분석을 수행하였다. 본 균주는 5,308,062 bp, G+C 비율 35.32%의 염색체와 308,946 bp, 32.23% G+C 함량이 포함된 plasmid를 지닌 것으로 확인되었다. 염색체와 plasmid DNA에 예측된 유전자의 총 수는 5,683개의 단백질 코딩유전자와 107개 tRNA 그리고 42개의 rRNA였다.

• Journal of Microbiology and Biotechnology
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• 제18권11호
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• pp.1836-1840
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• 2008

Bacteriocins are antimicrobial peptides that are produced by bacteria and toxic to bacterial strains closely related to the producer strain. It has previously been reported that Bacillus thuringiensis strain NEB17 and Bacillus thuringiensis subsp. kurstaki BUPM4 produce the bacteriocins thuricin 17 (3,162 Da) and bacthuricin F4 (3,160.05 Da), respectively. Here, we demonstrate that these bacteriocins have functional similarities and show a similar spectrum of antimicrobial activities against indicator strains. We also studied the effects of sterilization methods on the recovery and biological activities of these bacteriocins. They were completely degraded by autoclaving and the two were similarly affected by the tested filter membranes. Polyvinylidene fluoride (PVDF), polyestersulfone (PES), and cellulose acetate (CA) are suitable for filter sterilization of these bacteriocins. The two bacteriocins were stable across a range of storage conditions. These data will facilitate their utilization in food preservation or agricultural applications.

• International Journal of Industrial Entomology and Biomaterials
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• 제2권2호
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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.

• 한국미생물·생명공학회지
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• 제17권1호
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• pp.81-83
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• 1989

The flagella antigenic variation of nine Bacillus thuringiensis serovar kurstaki temperature-sensitive mutants grown at the permissive temperature (3$0^{\circ}C$) was detected by a serological agglutination between H-antigen and antiserum. The flagella antigens were injected to rabbits to prepared their antisera, and then their homologous and heterologous titers of the antisera were measured. The homologous titers were ranged from 1:6,400 to 1:12,800, but the heterologous titers were very low. The H-antigen of the wild type strain was not agglutinated to 4 heterologous antisera, ts-U23 not to 7, ts-U3l not 5, ts-U32 not to 4, ts-U33 not to 7, ts-U7l not to 4, ts-U73 not to 6, ts-U74 not to 6, ts-U91 not to 4 and ts-U603 not to 4 antisera.

• 한국미생물·생명공학회지
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• 제22권3호
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• pp.227-232
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• 1994

In order to microbially control root-knot nematode(Meloidogyne incognita) in tomato, a strain BtTH109 of Bacillus thuringiensis producing root-knot nematocidal toxin was isolated. The strain BtTH109 was identified B. thuringiensis subsp. indiana(serotype 16) based on flagella antigenicity, biochemical properties, and morphological charcateristics. The strain BtTH109 have extracellularly produced a root-knot nematocidal toxin, which was very toxic against not only egghatch but also the 2nd-nematode larva of root-knot nematode in vitro.

• 농약과학회지
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• 제15권2호
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• pp.166-176
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• 2011

제주도의 녹차 밭에 서식하는 딱정벌레목인 청동풍뎅이 (Anomala albopilosa)의 사체와 녹차 밭 토양에서 분리한 Bacillus thuringiensis CAB530 균주의 생물효과를 검토하였다. 이 균주는 몇 종류의 해충에 대한 살충활성에서 난방제 농업해충 가운데 하나인 파밤나방에 높은 효과를 나타냈다. 파밤나방 2령 유충에 대한 살충활성 검정에서 CAB530 균주는 $LC_{50}$값이 $1.49{\times}10^4$(cfu/$m{\ell}$)으로 고활성을 보였다. 이 균주가 생산하는 살충성 독소단백질의 SDS-PAGE에서는 파밤나방에 살충활성이 있는 기존의 B. thuringiensis subsp. kurstaki와 비슷한 130kDa의 밴드를 나타내었다. 또한 파밤나방 중장액으로 반응을 시킨 후에 약 65kDa의 활성 독성단백질을 확인할 수 있었다 PCR수행에서 CAB530 균주는 cry1Aa, cry1Ab, cry1C, cry1D, cry1F 그리고 cry1I등 6 개의 유전자가 존재하는 것으로 밝혀졌으며, B. thuringiensis subsp. kurstala기준 균주와 차이가 있었다. 딱정벌레목에서 분리 선발한 B. thuringiensis CAB530균주는 crystal의 형태와 SDS-PAGE의 결과는 B. thuringiensis subsp. kurstaki와 유사하게 나타났지만, 살충활성 검정과 PCR product 전기영동 결과는 B thuringiensis subsp. aizawai와 유사하게 나타났다.

• 미생물학회지
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• 제23권4호
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• pp.271-281
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• 1985

Bacillus thuringiensis var. thuringiensis produces an extracellular insecticidal thermostable .betha.-exotoxin, which was purified through microfiltering, barium precipitation, charcoal absorption chromatography, ion exchange column chromatography and gel filtration. The exotoxin in each purification step was detedted by thin layer chromatography, high pressure liquid chromatography and paper electrophoresis with efficient results. The exotoxin productivity on time course was checked by spectrophotometric absorbance at 258nm with the result that the exotoxin was initially produced in 6 hour culture and reached maximum value in 36 hour culture. Anti-bacterial effect test on Micrococcus flava was applied as toxicity test. The results showed that frowth inhibition of M. flava could be shown in plate assay of cell free filtered supernatant, alkaline eluant from charcoal and purified exotosin obtained from gel filtration column chromatography on Sephadex G-10 appeared to be 740. Heat stability of the exotoxin was confirmed through autoclaving twice.

• Journal of Biosystems Engineering
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• 제43권4호
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• pp.394-400
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• 2018

Purpose: This research evaluated five types of nanoparticles to develop a surface-enhanced Raman spectroscopy (SERS) method for the rapid detection of two Bacillus species (Bacillus cereus and Bacillus thuringiensis) that are commonly found on fresh produce, which can cause food poisoning. Methods: Bacterial concentrations were adjusted to a constant turbidity, and a total of $30{\mu}L$ of each Bacillus cell suspension was prepared for each nanoparticle. A point-scan Raman system with laser light source of wavelength 785 nm was used to obtain SERS data. Results: There was no qualitative difference in the SERS data of B. cereus and B. thuringiensis for any of the five nanoparticles. Three gold nanoparticles, stabilized in either citrate buffer or ethanol, showed subtle differences in Raman intensities of two Bacillus species at $877.7cm^{-1}$. Conclusions: Among the three types of nanoparticles, the gold nanoparticles stabilized in citrate buffer showed the lowest standard deviation, followed by gold nanoparticles stabilized in ethanol. This result supports the potential application of gold nanoparticles for SERS-based detection of B. cereus and B. thuringiensis.

• 한국응용곤충학회지
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• 제35권1호
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• pp.66-73
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• 1996

Bacillus thuringiensis subsp. morrisoni PG-14의 cryIVD 유전자를 포함하고 있는 발현벡터 pCYASK5-1을 제작하여 Synechocystis PCCC6803에 형질전환시킨 후, 학질모기(Anopheles sinensis)유충에 대한 독성을 검정하였다. Kanamycin이 포함된 BG-11배지에서 선발된 형질전환체의 cryIVD 유전자 발현은 SDS-PAGE와 West-ern blot분석으로 확인하였다. 형질전환체는 A. sinensis 유충에 대해 높은 독성을 나타내었으며, 성장은 야생주인 Synechocystis Pccc6803과 유사하였다. 또 수심에 따른 형질전환체의 분포도 조사에서 전체적으로 살충농도의 세포수로 분포함을 확인하였다. 이러한 결과들은 B. thuringiensis subsp. morrisoni PG-14의 cryIVD 유전자로 형질전환된 Synechocystis PCCC6803이 모기유충 방제에 효율적으로 이용될 수 있음을 나타내었다.