Research Support, Non-U.S. Gov't
- Benzaldehyde as an insecticidal, antimicrobial, and antioxidant compound produced by Photorhabdus temperata M1021
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Ihsan Ullah , Abdul Latif Khan , Liaqat Ali , Abdur Rahim Khan , Muhammad Waqas , Javid Hussain , In-Jung Lee , Jae-Ho Shin
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J. Microbiol. 2015;53(2):127-133. Published online January 28, 2015
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DOI: https://doi.org/10.1007/s12275-015-4632-4
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Abstract
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The Photorhabdus temperata M1021 secretes toxic compounds
that kill their insect hosts by arresting immune
responses. Present study was aimed to purify the insecticidal
and antimicrobial compound(s) from the culture extract of
P. temperata M1021 through bioassay guided fractionation.
An ethyl acetate (EtOAc) extract of the P. temperata M1021
exhibited 100% mortality in Galleria mellonella larvae within
72 h. In addition, EtOAc extract and bioactive compound 1
purified form the extract through to column chromatography,
showed phenol oxidase inhibition up to 60% and
80% respectively. The analysis of 1H and 13C NMR spectra
revealed the identity of pure compound as "benzaldehyde".
The benzaldehyde showed insecticidal activity against G.
mellonella in a dose-dependent manner and 100% insect
mortality was observed at 108 h after injection of 8 mM
benzaldehyde. In a PO inhibition assay, 4, 6, and 8 mM concentrations
of benzaldehyde were found to inhibit PO activity
about 15%, 42%, and 80% respectively. In addition,
nodule formation was significantly (P < 0.05) inhibited by 4,
6, and 8 mM of benzaldehyde as compare to control. Moreover,
benzaldehyde was found to have great antioxidant activity
and maximum antioxidant activity was 52.9% at 8 mM
benzaldehyde as compare to control. Antimicrobial activity
was assessed by MIC values ranged from 6 mM 10 mM for
bacterial strains and 8 mM to 10 mM for fungal strains. The
results
suggest that benzaldehyde could be applicable for
developing novel insecticide for agriculture use.
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Journal Article
- Aeration Effects on Metabolic Events during Sporulation of Bacillus thuringiensis
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Mohammad H. Sarrafzadeh , Sabine Schorr-Galindo , Hyun-Joon La , Hee-Mock Oh
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J. Microbiol. 2014;52(7):597-603. Published online June 28, 2014
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DOI: https://doi.org/10.1007/s12275-014-3547-9
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The metabolism of Bacillus thuringiensis during its sporulation process was investigated under different concentrations of oxygen. At the beginning of sporulation, the aeration conditions were regulated to obtain different oxygen transfer rates (OTR) in four separate fermentations, representing interrupted, limited, non-limited, and saturated oxygenation, respectively. A higher OTR resulted in a higher pH, up to about 9 in the case of saturated oxygenation, while the interrupted oxygenation resulted in a significantly acidic culture. In contrast, the absence of oxygen resulted in rapid sporangia lysis and caused acidification of the medium, indicating a distinctly different sporangia composition and different metabolism. The bacterium also showed different CO2 production rates during sporulation, although amaximum point was observed in every case.With a higher OTR, the maximal value was observed after a longer time and at a lower value (40, 26, and 13 mmol/L/h for limited, non-limited, and saturated cases, respectively). Despite the exhaustion of glucose prior to the sporulation phase, the interrupted oxygenation resulted in acetate, lactate, and citrate in the medium with a maximum concentration of 4.8, 1.3, and 5.0 g/L, respectively. Notwithstanding, while the metabolic events differed visibly in the absence of oxygen, once sporulation was triggered, it was completed, even in the case of an interrupted oxygen supply.
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Research Support, Non-U.S. Gov'ts
- Comparative Phylogenetic Relationships and Genetic Structure of the Caterpillar Fungus Ophiocordyceps sinensis and Its Host Insects Inferred from Multiple Gene Sequences
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Qing-Mei Quan , Qing-Xia Wang , Xue-Li Zhou , Shan Li , Xiao-Ling Yang , Yun-Guo Zhu , Zhou Cheng
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J. Microbiol. 2014;52(2):99-105. Published online February 1, 2014
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DOI: https://doi.org/10.1007/s12275-014-3391-y
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295
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Ophiocordyceps sinensis (Ascomycota: Ophiocordycipitaceae) is a native fungal parasite of Hepialidae caterpillars and one of the most economically important medicinal caterpillar fungi in China. However, little is known about the phylogenetic and evolutionary relationships between O. sinensis and its host insects. In this study, nuclear ITS and β-tubulin sequences from O. sinensis and mitochondrial COI, COII, and Cytb sequences from its hosts were analyzed across 33 populations sampled from five regions in China. Phylogenetically, both O. sinensis and its hosts were divided into three geographically correlated clades, and their phylogenies were congruent. Analysis of molecular variance and calculated coefficients of genetic differentiation revealed significant genetic divergence among the clades within both O. sinensis (FST=0.878, NST=0.842) and its hosts (FST=0.861, NST=0.816). Estimated gene flow was very low for O. sinensis (Nm=0.04) and the host insects (Nm=0.04) among these three clades. Mantel tests demonstrated a significant correlation (P<0.01) between the genetic distances for O. sinensis and its hosts, as well as a significant association (P<0.05) between geographic and genetic distances in both. The similar phylogenetic relationships, geographic distributions, and genetic structure and differentiation between O. sinensis and its hosts imply that they have coevolved.
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- Sequential Immunosuppressive Activities of Bacterial Secondary Metabolites from the Entomopahogenic Bacterium Xenorhabdus nematophila
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Seonghyeon Eom , Youngjin Park , Yonggyun Kim
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J. Microbiol. 2014;52(2):161-168. Published online February 1, 2014
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DOI: https://doi.org/10.1007/s12275-014-3251-9
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300
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The entomopathogenic bacterium Xenorhabdus nematophila secretes at least eight bacterial metabolites that play crucial roles suppressing target insect immune responses by inhibiting eicosanoid biosynthesis. We analyzed sequential changes in bacterial metabolite production during bacterial growth and analyzed their individual immunosuppressive activities against the insect host, Spodoptera exigua. X. nematophila exhibited a typical bacterial growth pattern in both insect host and culture medium, and eight metabolites were secreted at different time points. At the early growth phase (6–12 h), Ac-FGV and PHPP were detected in significant amounts in the culture broth. At this early phase, both Ac-FGV (18 μg/ml) and oxindole (110 μg/ml) levels significantly inhibited phenoloxidase and phospholipase A2 activities in S. exigua hemolymph. At the late growth phase (12–36 h), all eight metabolites were detected at significant levels (10–140 μg/ml) in the culture broth and were sufficient to induce hemocyte toxicity. These results suggest that X. nematophila sequentially produces immunosuppressive metabolites that might sequentially and cooperatively inhibit different steps of insect immune responses.
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Shabbir Ahmed, Yonggyun Kim
Journal of Invertebrate Pathology.2018; 157: 136. CrossRef - Challenging the larvae of Helicoverpa armigera and assessing the immune responses to nematode-bacterium complex
Istkhar, Ashok Kumar Chaubey
Phytoparasitica.2018; 46(1): 75. CrossRef - Refining the Natural Product Repertoire in Entomopathogenic Bacteria
Nicholas Jay Tobias, Yi-Ming Shi, Helge B. Bode
Trends in Microbiology.2018; 26(10): 833. CrossRef - Role of Endosymbionts in Insect–Parasitic Nematode Interactions
Ioannis Eleftherianos, Shruti Yadav, Eric Kenney, Dustin Cooper, Yaprak Ozakman, Jelena Patrnogic
Trends in Parasitology.2018; 34(5): 430. CrossRef - Eicosanoid mediation of immune responses at early bacterial infection stage and its inhibition by Photorhabdus temperata subsp. temperata, an entomopathogenic bacterium
Hyoil Kim, Duyeol Choi, Jihyeon Jung, Yonggyun Kim
Archives of Insect Biochemistry and Physiology.2018;[Epub] CrossRef - Identification and bacterial characteristics of Xenorhabdus hominickii ANU101 from an entomopathogenic nematode, Steinernema monticolum
Youngjin Park, Sangjin Kang, Md. Sadekuzzaman, Hyeonghwan Kim, Jin-Kyo Jung, Yonggyun Kim
Journal of Invertebrate Pathology.2017; 144: 74. CrossRef - The Global Transcription Factor Lrp Is both Essential for and Inhibitory to Xenorhabdus nematophila Insecticidal Activity
Ángel M. Casanova-Torres, Upasana Shokal, Neta Morag, Ioannis Eleftherianos, Heidi Goodrich-Blair, Harold L. Drake
Applied and Environmental Microbiology.2017;[Epub] CrossRef - Effects of an entomopathogen nematode on the immune response of the insect pest red palm weevil: Focus on the host antimicrobial response
Simona Binda-Rossetti, Maristella Mastore, Marina Protasoni, Maurizio F. Brivio
Journal of Invertebrate Pathology.2016; 133: 110. CrossRef - A Mixture ofBacillus thuringiensissubsp.israelensisWithXenorhabdus nematophila-Cultured Broth Enhances Toxicity Against MosquitoesAedes albopictusandCulex pipiens pallens(Diptera: Culicidae)
Youngjin Park, Jin Kyo Jung, Yonggyun Kim
Journal of Economic Entomology.2016; 109(3): 1086. CrossRef - Microbiology of sugar‐rich environments: diversity, ecology and system constraints
Bart Lievens, John E. Hallsworth, Maria I. Pozo, Zouhaier Ben Belgacem, Andrew Stevenson, Kris A. Willems, Hans Jacquemyn
Environmental Microbiology.2015; 17(2): 278. CrossRef - Comparison of Xenorhabdus bovienii bacterial strain genomes reveals diversity in symbiotic functions
Kristen E. Murfin, Amy C. Whooley, Jonathan L. Klassen, Heidi Goodrich-Blair
BMC Genomics.2015;[Epub] CrossRef - Entomopathogenic bacterium, Xenorhabdus nematophila and Photorhabdus luminescens, enhances Bacillus thuringiensis Cry4Ba toxicity against yellow fever mosquito, Aedes aegypti (Diptera: Culicidae)
Youngjin Park
Journal of Asia-Pacific Entomology.2015; 18(3): 459. CrossRef - A Technique to Enhance Insecticidal Efficacy Using Bt Cry Toxin Mixture and Eicosanoid Biosynthesis Inhibitor
Seonghyeon Eom, Youngjin Park, Yonggyun Kim
The Korean Journal of Pesticide Science.2015; 19(3): 301. CrossRef - Anopheles gambiae eicosanoids modulate Plasmodium berghei survival from oocyst to salivary gland invasion
Susana Ramos, Ana Custódio, Henrique Silveira
Memórias do Instituto Oswaldo Cruz.2014; 109(5): 668. CrossRef - Integrin β subunit and its RNA interference in immune and developmental processes of the Oriental tobacco budworm, Helicoverpa assulta
Youngjin Park, Seung-Joon Ahn, Heiko Vogel, Yonggyun Kim
Developmental & Comparative Immunology.2014; 47(1): 59. CrossRef - A Technique to Enhance Bacillus thuringiensis Spectrum and Control Efficacy Using Cry Toxin Mixture and Immunosuppressant
Seonghyeon Eom, Youngjin Park, Yonggyun Kim
The Korean Journal of Pesticide Science.2014; 18(3): 181. CrossRef - Effect of Cellular Phospholipase A2Inhibition on Enhancement of Bt Insecticidal Activity
Seonghyeon Eom, Jiyeong Park, Kunwoo Kim, Yonggyun Kim
Korean journal of applied entomology.2014; 53(3): 271. CrossRef
- NOTE] Construction and Characterisation of an Antifungal Recombinant Bacillus thuringiensis with an Expanded Host Spectrum
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Qin Liu , Jong Yul Roh , Yong Wang , Jae Young Choi , Xue Ying Tao , Jae Su Kim , Yeon Ho Je
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J. Microbiol. 2012;50(5):874-877. Published online November 4, 2012
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DOI: https://doi.org/10.1007/s12275-012-2201-7
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A novel antifungal Bacillus thuringiensis strain 19–22, ssp. kurstaki (H3a3b3c), was characterised. This strain included cry1Aa, cry1Ab, cry1Ac, and cry1D, which have high insecticidal activities against lepidopteran larvae other than Spodoptera exigua. To expand the host spectrum, a cry1E gene whose product is active against S. exigua was introduced into the isolate. The transformant successfully expressed the Cry1E protein without any loss of its original antifungal activities. These results indicate that this recombinant strain exhibits dual activities and may be used as an integrated control agent to control plant diseases and insect pests.
- Evaluation of Insecticidal Activity of a Bacterial Strain, Serratia sp. EML-SE1 against Diamondback Moth
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Hyung Uk Jeong , Hye Yeon Mun , Hyung Keun Oh , Seung Bum Kim , Kwang Yeol Yang , Iksoo Kim , Hyang Burm Lee
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J. Microbiol. 2010;48(4):541-545. Published online August 20, 2010
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DOI: https://doi.org/10.1007/s12275-010-0221-9
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To identify novel bioinsecticidal agents, a bacterial strain, Serratia sp. EML-SE1, was isolated from a dead larva of the lepidopteran diamondback moth (Plutella xylostella) collected from a cabbage field in Korea. In this study, the insecticidal activity of liquid cultures in Luria-Bertani broth (LBB) and nutrient broth (NB) of a bacterial strain, Serratia sp. EML-SE1 against thirty 3rd and 4th instar larvae of the diamondback moth was investigated on a Chinese cabbage leaf housed in a round plastic cage (Ø 10×6 cm). 72 h after spraying the cabbage leaf with LBB and NB cultures containing the bacterial strain, the mortalities of the larvae were determined to be 91.7% and 88.3%, respectively. In addition, the insecticidal activity on potted cabbage containing 14 leaves in a growth cage (165×83×124 cm) was found to be similar to that of the plastic cage experiment. The results of this study provided valuable information on the insecticidal activity of the liquid culture of a Serratia species against the diamondback moth.
- NOTE] Biosynthesis of Poly(3-Hydroxybutyrate-co-3-Hydroxyvalerate) Copolyesters with a High Molar Fraction of 3-Hydroxyvalerate by an Insect-Symbiotic Burkholderia sp. IS-01
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Do Young Kim , Doo-Sang Park , Soon Bum Kwon , Moon Gyu Chung , Kyung Sook Bae , Ho-Yong Park , Young Ha Rhee
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J. Microbiol. 2009;47(5):651-656. Published online October 24, 2009
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DOI: https://doi.org/10.1007/s12275-009-0109-7
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Burkholderia sp. IS-01 capable of biosynthesizing poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [poly(3HB- co-3HV)] copolyesters with a high molar fraction of 3HV was isolated from the gut of the adult longicorn beetle, Moechotypa diphysis. The strain IS-01 was relatively tolerant to high concentrations of levulinic acid and accumulated a poly(13.5 mol% 3HB-co-86.5 mol% 3HV) copolyester when cultivated on a mixture of gluconate (20 g/L) and levulinic acid (12.5 g/L). In this case, the content of the copolyester in the cells was approximately 60.0%. The compositions of the copolyesters were easily regulated by altering the molar ratio of gluconate and levulinic acid in the medium. The organism was found to possess a class I PHA synthase (PhaC) gene (1,881 bp) that encodes a protein with a deduced molecular mass of 68,538 Da that consists of 626 amino acids. The PhaC of this organism was most similar to that of B. cenocepacia PC184 (92% similarity).
- Characterization of an Extracellular Lipase in Burkholderia sp. HY-10 Isolated from a Longicorn Beetle
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Doo-Sang Park , Hyun-Woo Oh , Sun-Yeon Heo , Woo-Jin Jeong , Dong Ha Shin , Kyung Sook Bae , Ho-Young Park
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J. Microbiol. 2007;45(5):409-417.
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DOI: https://doi.org/2596 [pii]
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Burkholderia sp. HY-10 isolated from the digestive tracts of the longicorn beetle, Prionus insularis, produced an extracellular lipase with a molecular weight of 33.5 kDa estimated by SDS-PAGE. The lipase was purified from the culture supernatant to near electrophoretic homogenity by a one-step adsorption-desorption procedure using a polypropylene matrix followed by a concentration step. The purified lipase exhibited highest activities at pH 8.5 and 60°C. A broad range of lipase substrates, from C4 to C18 ρ-nitrophenyl esters, were hydrolyzed efficiently by the lipase. The most efficient substrate was ρ-nitrophenyl caproate (C6). A 2485 bp DNA fragment was isolated by PCR amplification and chromosomal walking which encoded two polypeptides of 364 and 346 amino acids, identified as a lipase and a lipase foldase, respectively. The N-terminal amino acid sequence of the purified lipase and nucleotide sequence analysis predicted that the precursor lipase was proteolytically modified through the secretion step and produced a catalytically active 33.5 kDa protein. The deduced amino acid sequence for the lipase shared extensive similarity with those of the lipase family I.2 of lipases from other bacteria. The deduced amino acid sequence contained two Cystein residues forming a disulfide bond in the molecule and three, well-conserved amino acid residues, Ser131, His330, and Asp308, which composed the catalytic triad of the enzyme.
- Propagation of Bombyx mori Nucleopolyhedrovirus in Nonpermissive Insect Cell Lines
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Soo-Dong Woo , Jong Yul Roh , Jae Young Choi , Byung Rae Jin
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J. Microbiol. 2007;45(2):133-138.
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DOI: https://doi.org/2522 [pii]
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This study addresses the susceptibility of Spodoptera frugiperda (Sf9 and Sf21), Trichoplusia ni (Hi5), and S. exigua (Se301) cells to the Bombyx mori nucleopolyhedrovirus (BmNPV). Although these cells have classically been considered nonpermissive to BmNPV, the cytopathic effect, an increase in viral yield, and viral DNA synthesis by BmNPV were observed in Sf9, Sf21, and Hi5 cells, but not in Se301 cells. Very late gene expression by BmNPV in these cell lines was also detected via β-galactosidase expression under the control of the polyhedrin promoter. Sf9 cells were most susceptible to BmNPV in all respects, followed by Sf21 and Hi5 cells in decreasing order, while the Se301 cells evidenced no distinct viral replication. This particular difference in viral susceptibility in each of the cell lines can be utilized for our understanding of the mechanisms underlying the host specificity of NPVs.
- Investigations on Bacteria as a Potential Biological Control Agent of Summer Chafer, Amphimallon solstitiale L. (Coleoptera: Scarabaeidae)
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Kazlm Sezen , Ismail Demir , Hatice Katl , Zihni Demirbag
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J. Microbiol. 2005;43(5):463-468.
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DOI: https://doi.org/2274 [pii]
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Studying the bacteria of hazardous insects allows the opportunity to find potentially better biological control agents. Therefore, in this study, bacteria from summer chafer (Amphimallon solstitiale L., Coleoptera: Scarabaeidae) we isolated and identified the insecticidal effects of bacteria isolated from A. solstitiale and Melolontha melolontha L. (common cockchafer, Coleoptera: Scarabaeidae) and the mixtures of these bacterial isolates were investigated on A. solstitiale larvae. Crystals from Bacillus sp. isolated from M. melolontha were also purified, and tested against the second and third-stage larvae of A. solstitiale. The bacterial isolates of A. solstitiale were identified as Pseudomonas sp., Pseudomonas sp., Bacillus cereus and Micrococcus luteus, based on their morphology, spore formation, nutritional features, and physiological and biochemical characteristics. The insecticidal effects of the bacterial isolates determined on the larvae of A. solstitiale were 90% with B. cereus isolated from A. solstitiale, and 75% with B. cereus, B. sphaericus and B. thuringiensis isolated from M. melolontha within ten days. The highest insecticidal effects of the mixed infections on the larvae of A. solstitiale were 100% both with B. cereus+B. sphaericus and with B. cereus+B. thuringiensis. In the crystal protein bioassays, the highest insecticidal effect was 65% with crystals of B. thuringiensis and B. sphaericus isolated from M. melolontha within seven days. Finally, our results showed that the mixed infections could be utilized as microbial control agents, as they have a 100% insecticidal effect on the larvae of A. solstitiale.
- Overexpression of Insecticidal Protein Gene of Bacillus thuringiensis var. kurstaki HD1
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Hwang, Sung Hei , Yoo, Kwan Hee , Moon, Eui Sik , Cha Soung Chul , Lee, Hyung Hoan
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J. Microbiol. 1998;36(4):289-295.
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The Insecticidal protein (ICP) gene from Bacillus thuringiensis var, kurstaki HD-1 was cloned in pBluescript SK(+) vector and charicterized by overexpression in Escherichia coli XL1-blue. Total plasmids in the B. thuringiensis were isolated and digested with restriction enzyme BamHI. Then, southern blot was performed with a probe to locate the gene in the fragments. The hybridized 3.8 kb NdeI DNA fragment was cloned into the SmaI site of pBluescript SK(+) and named pHLN1-80 in forward orientation to the lacZ gene promoter and pHLN2-80 in reverse orientation to the lacZ gene promoter. Determination of 153 bp nucleotide sequence of 5'-end of the NdeI fragment in the pHLN1-80 clone revealed that there are-80 bp region of the ICP gene promoter and +73 bp region of the ICP gene at the 5'end of the ICP gene. In addition, the-80 bp promoter of the ICP gene contained transcription initiation point G at-77 bp point and BtI promoter and Shine-Dalgarno sequence at-14 to-4 bp region. The two clones showed strong insecticidal activity against 3rd the instar Bompyx mori larvae. SDS-PAGE analysis revealed that the pHLN2-80 clone clearly produces distinguishable amount (27 times more) of the 130 kDa ICP band and 100 times the insecticidal activity than that of the clone pHLN1-80. These marked differences in production and toxicity due to different orientations of the gene in the vectior provide us valuable points for further study on the ICP gene transcription at the molecular level.