• 제목/요약/키워드: bipyramidal crystals

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국내 식물시료에서 분리한 Bacillus thuringiensis 균주의 다양성

  • 박승환;구본탁;신병식;최수근;정영미;반재구;김정일
    • 한국미생물·생명공학회지
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    • 제25권2호
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    • pp.159-165
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    • 1997
  • We collected 3,237 plant samples, mainly leaves of various trees, from many provinces in Korea and a total of 1,925 Bacillus thuringiensis isolates were obtained and characterized. The isolates were characterized in terms of crystal morphology, PAGE pattern of the toxin proteins, plasmids pattern, biochemical characteristics, and bioassay. The microscopic observation showed that 49.1% of the isolates have bipyramidal shape crystals, 7.1% of spherical shape crystals, 1.4% of rhomboidal shape crystals, and others have small or amorphous inclusions. The insecticidal activities of the spore-crystal mixtures of isolates were tested against Plutella xylostella, Bombyx mori, Culex pipiens, and Agelastica coerulea. Bioassay showed that 51.3% of the isolates were shown to be active; lepidopteran-specific (44.8%), dipteran-specific(4.9%) and coleopteran-specific (1.6%). The remainder(48.8%) did not show any activity against the insects we tested. Interestingly though, some of these non-active isolates were shown to have bipyramidal crystals. By serotyping 22 isolates of our collection, we found that there are various kind of subspecies such as aizawai, amagiens, canadensis, darmstadiensis, galleriae, finitimus, kurstaki, morrisoni and neoleonensis, and three isolates have been classified into a new serotype, H49, and one of them, the type strain, named subsp. muju. From this study it was found that phylloplane is a good source for the isolation of Bacillius thuringiensis, and Bacillus thuringiensis is distributed widely in Korea.

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Characterization of bacillus thuringiensis isolates form soil in wonju area

  • Yoo, Kwan-Hee;Kim, Soo-Young;Ho, Kang-Min;Cho, Myung-Hwan;Lee, Hyung-Hoan
    • Journal of Microbiology
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    • 제34권4호
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    • pp.370-373
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    • 1996
  • Three strains (KW-1, KW-14, KW-15) of Bacillus thuringiensis were isolated from soil in Wonju area and characterized. The three strains produced parasporal inclusion bodies (crystals) and spores in their cells. The KW-1 strain produces spherical crystals. The crystals of strain KW-14 are bipyramidal crystal. The KW-15 strain harbors irregular crystals. Only minor biochemical characteristics of the three isolates were different and distinctive, however general characteristics were similar to the known serotypes of B. thuringiensis. Three strains were resistant to penicilin G, oxacillin and cephalothin. Three strains were highly toxic to Bombyx mori larvae, but not to the Culex pipiens larvae.

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Chemistry of Ruthenium Hydridonitrosyl Complexes Containing Chelating Triphosphines I-Structures of RuH(NO)$P_3$ ($P_3$ : Chelating Triphosphines)

  • Ik Mo Lee;Devon W. Mee;Judith Gallucci
    • Bulletin of the Korean Chemical Society
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    • 제13권5호
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    • pp.491-498
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    • 1992
  • Chelating triphosphines were applied to freeze the fluxionality and to minimize the number of isomers found in the monophosphine analogues and this technique was proved to be useful. RuH(NO)$P_3$($P_3$; Cyttp, ttp and etp) complexes were characterized to have similar trigonal bipyramidal structures with linear NO groups. Cyttp prefers to have a meridional geometry while etp prefers a facial one and ttp complexes are mixture of these two isomers. The crystal structure of RuH(NO)(Cyttp) has been determined to have a distorted trigonal bipyramidal structure with a linear NO in the equatorial plane. The crystals are orthorhombic, space group $P_{nma}$, with unit cell dimensions a = 16.356(2), b = 20.474(2), c = 10.915(l) ${\AA}$, V = 3655 ${\AA}^3$, Z = 4, R = 0.035 and $R_w$ = 0.034 for the 2900 intensities with $F_o^2 >3{\sigma}(F_o^2)$ and the 208 variables.

Insecticidal Characterization of Thirteen Bacillus thuringiensis Isolates from Soil (III)

  • Lee, Hyung H.;Lee, Kwang Y.;Kim, Tae-J;Sun B. Sim;Joong G. Cho;Sun I. Kwon
    • 미생물학회지
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    • 제30권6호
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    • pp.438-443
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    • 1992
  • Thirteen strains of Bacillus thuringiensis were isolated from soil in Korea and characterized. The all strains produced parasporal crystals and spores in their cells. Two strains had bipyramidal crystals, seven strains contianed round ones and four strains had unregular ones. Only minor biochemical characteristics of the thirteen isolates were different and distinctive, however general characteristics were similar to the known serotypes of B. thuringiensis. Two strains were resistant to ampicilin. Three strains were resistant to bacitracin, six strains were resistant to cephalothin, two strains were resistant to colistin, HL-68 strain was resistant to gentamycin, HL-67 strain was resistant to kanamycin and HL-71 was resistant to tetracycline. Two strains were resistant to penicillin G. Four strains were toxic to Bombyx mori larvae and eleven strains were toxic to Culex pipiens larvae.

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Mössbauer Study of the Dynamics in BaFe12O19 Single Crystals

  • Choi, J.W.;Sur, J.C.;Lim, Jung-Tae;Kim, Chin-Mo;Kim, Chul-Sung
    • Journal of Magnetics
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    • 제17권1호
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    • pp.6-8
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    • 2012
  • M$\ddot{o}$ssbauer spectra of hexagonal $BaFe_{12}O_{19}$ single crystals were studied at various temperatures (4-300 K). It was found that the spin states in Fe atoms were parallel to the ${\gamma}$-ray's direction into a single crystal along the caxis. The location of the Fe ion in the 2b site is unusual in an oxide structure and has strong anisotropic lattice vibrations. Moreover, at room temperature, the zero absorption lines of the Fe ions at the 2b site were observed due to fast diffusion motion in a double well atomic potential. The two Fe ions of the single crystal mainly enter into the sites in the mirror plane of the trigonalbipyramidal structure.

Electron Microscopic Evidence of Paraporal Crystal Inclusion Biogenesis in Bacillus sphaericus Strain 1593

  • Lee, Young-Ju;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • 제11권6호
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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).

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Spacing Effect of the Intervening Sequences between Ribosome Binding Site and the Initiation Codon on Expression of Bacillus thuringiensis $\delta$-Endotoxin

  • Roh, Jong-Yul;Li, Ming-Shun;Chang, Jin-Hee;Park, Jae-Young;Shim, Hee-Jin;Woo, Soo-Dong;Boo, Kyung-Saeng;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • 제6권1호
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    • pp.81-85
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    • 2003
  • To verify importance of the intervening sequence between the ribosome binding site (RBS) and the initiation codon for expression of Bacillus thuringiensis $\delta$-endotoxin, the pProMu, containing SphI and NcoIsites between RBS and the initiation codon of the cry1Ac gene, and the deletion derivatives of pProMu were constructed and transformed into the B. thuringiensis subsp. kurstaki $Cry^{-B}$ strain. The pProMu-ΔSphIhad identical six bases of intervening sequence to pProAc though the arrangement of sequence was different. Other mutants containing pProMu had 1 or 10 or 14 bases between RBS and the initiation codon. Among deletion mutants, only ProMu-ΔSphI/CB only produced 130 kDa typical bipyramidal crystals like those seen for ProAc/CB. However, ProMu/CB, $ProMu-{\Delta}NcoI$, and ProMu-ΔSphI+NcoIdid not produce Cry1Ac crystals. In conclusion, the results suggest that 6-base intervening sequence was important for expression of cry1-type class gene. Furthermore, spacing effect of the intervening sequences may play an important role in expression of individual crystal proteins in B. thuringiensis without doubt.

Bacillus thuringiensis 내에서 안정한 벡타를 이용한 cry1C 유전자의 발현

  • 최수근;오근희;김정일;박승환
    • 한국미생물·생명공학회지
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    • 제25권6호
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    • pp.566-570
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    • 1997
  • During sporulation, Bacillus thuringiensis strains produce crystals consist of toxin proteins highly specific against insect pests. Their host specificities are desirable from a standpoint of environmental safety, but also limit market potential. Thus, development of improved Bacillus thuringiensis strains having broad host spectrum will contribute to increase its use. For the construction of Bacillus thuringiensis strain having broad host spectrum, we cloned cry1C gene encoding a toxin protein highly toxic against Spodoptera exigua from a B. thuringiensis isolate and constructed two recombinant plasmids, pUBClC and plC60. The plasmid PUBC1C has a replication origin of the natural plasmid pBC16 from B. cereus which is closely related species to B. thuringiensis, and the pBC16 was known to be replicated by rolling-circle mechanism. The plasmid pIC60 has a replication origin of a resident 60 MDa plasmid from B. thuringiensis subsp. kurstaki HD263, and it is believed that the pIC60 is replicated in a theta mode. The two plasmids were introduced into B. thuringiensis subsp. kurstaki cryB strain, and the transformed strains produced well-shaped bipyramidal crystals. We confirmed the expression of the cry1C gene by SDS-PAGE, and Western blotting. By investigating the segregational stability, it was found that the plasmid pIC60 is more stable than the pUBC1C.

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새로 분리한 Bacillus thuringiensis NT0423균주의 내독소 단백질에 대한 이중 특이성 (Dual specificity of $\delta$-endotoxins produced by newly isolated Bacillus thuringiensis NT0423)

  • 김호산;박현우;김상현;유용만;서숙재;강석권
    • 한국응용곤충학회지
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    • 제32권4호
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    • pp.426-432
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    • 1993
  • 경기도 일원의 양잠 농가의 먼지에서 채취한 45개의 샘플중에서 내독소 단백질 결정체를 생산하는 13개의 Bacillus thuringiensis를 분리하였다. 이 중 2개 균주는 파리목에 독성을 나타냈으며, 특히 독성검정에서 NT0423균주의 $LC_{50}$수치는 나비목의 배추좀나방이 최소 1.30$\times$$10^{6}$ CFU/ml이며, 파리목인 빨간 집모기에는 2.88$\times$$10^{5}$ CFU/ml로 나타났다. 새로 분리된 NT0423균주가 생산하는 내독소 단백질 결정체는 주사 전자현미경사진에서 전형적인 이중 피라미드모양을 보였다. 그리고 이 내독소 단백질 결정체의 SDS-PAGE 분석에서는 주요한 130kDa의 polypeptide을 나타내었다. 또한 NT0423균주의 총 플라스미드 DNA분석에서는 9개의 플라스미드를 갖고 있어 기존의 유사한 독성을 나타내는 균주들과 다른 패턴을 보였다.

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A Highly Pathogenic Strain of Bacillus thuringiensis serovar kurstaki in Lepidopteran Pests

  • Kati, Hatice;Sezen, Kazim;Nalcacioglu, Remziye;Demirbag, Zihni
    • Journal of Microbiology
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    • 제45권6호
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    • pp.553-557
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    • 2007
  • In order to detect and identify the most toxic Bacillus thuringiensis strains against pests, we isolated a B. thuringiensis strain (Bn1) from Balaninus nucum (Coleoptera: Curculionidae), the most damaging hazelnut pest. Bn1 was characterized via morphological, biochemical, and molecular techniques. The isolate was serotyped, and the results showed that Bn1 was the B. thuringiensis serovar, kurstaki (H3abc). The scanning electron microscopy indicated that Bn1 has crystals with cubic and bipyramidal shapes. The Polymerase Chain Reactions (PCRs) revealed the presence of the cry1 and cry2 genes. The presence of Cry1 and Cry2 proteins in the Bn1 isolate was confirmed via SDS-PAGE, at approximately 130 kDa and 65 kDa, respectively. The bioassays conducted to determine the insecticidal activity of the Bn1 isolate were conducted with four distinct insects, using spore-crystal mixtures. We noted that Bn1 has higher toxicity as compared with the standard B. thuringiensis subsp. kurstaki (HD-1). The highest observed mortality was 90% against Malacosoma neustria and Lymantria dispar larvae. Our results show that the B. thuringiensis isolate (Bn1) may prove valuable as a significant microbial control agent against lepidopteran pests.