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Research Support, Non-U.S. Gov't
Benzaldehyde as an insecticidal, antimicrobial, and antioxidant compound produced by Photorhabdus temperata M1021
Ihsan Ullah , Abdul Latif Khan , Liaqat Ali , Abdur Rahim Khan , Muhammad Waqas , Javid Hussain , In-Jung Lee , Jae-Ho Shin
J. Microbiol. 2015;53(2):127-133.   Published online January 28, 2015
DOI: https://doi.org/10.1007/s12275-015-4632-4
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AbstractAbstract
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
Mohammad H. Sarrafzadeh , Sabine Schorr-Galindo , Hyun-Joon La , Hee-Mock Oh
J. Microbiol. 2014;52(7):597-603.   Published online June 28, 2014
DOI: https://doi.org/10.1007/s12275-014-3547-9
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AbstractAbstract PDF
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
Qing-Mei Quan , Qing-Xia Wang , Xue-Li Zhou , Shan Li , Xiao-Ling Yang , Yun-Guo Zhu , Zhou Cheng
J. Microbiol. 2014;52(2):99-105.   Published online February 1, 2014
DOI: https://doi.org/10.1007/s12275-014-3391-y
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AbstractAbstract PDF
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
Seonghyeon Eom , Youngjin Park , Yonggyun Kim
J. Microbiol. 2014;52(2):161-168.   Published online February 1, 2014
DOI: https://doi.org/10.1007/s12275-014-3251-9
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AbstractAbstract PDF
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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    Ahmed Mohamed Abd El-Rah, Amr Mohamed Abdelazeem, Hanaa Elbrense, Santiago Vergara-Pi
    Pakistan Journal of Biological Sciences.2022; 25(7): 586.     CrossRef
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    Saqer S. Alotaibi, Hadeer Darwish, Sarah Alharthi, Akram Alghamdi, Ahmed Noureldeen, Ahmed M. Fallatah, András Fodor, Amal Al-Barty, Bander Albogami, Alaa Baazeem
    Agriculture.2021; 11(12): 1256.     CrossRef
  • Functional interaction of bacterial virulence factors of Xenorhabdus nematophila with a calcium-independent cytosolic PLA2 of Spodoptera exigua
    Vatanparast Mohammad, Yonggyun Kim
    Journal of Invertebrate Pathology.2020; 169: 107309.     CrossRef
  • Immunosuppressive Activities of Novel PLA2 Inhibitors from Xenorhabdus hominickii, an Entomopathogenic Bacterium
    Md. Mahi Imam Mollah, Aman Dekebo, Yonggyun Kim
    Insects.2020; 11(8): 505.     CrossRef
  • The great potential of entomopathogenic bacteria Xenorhabdus and Photorhabdus for mosquito control: a review
    Wellington Junior da Silva, Harry Luiz Pilz-Júnior, Ralf Heermann, Onilda Santos da Silva
    Parasites & Vectors.2020;[Epub]     CrossRef
  • Benzylideneacetone and other phenylethylamide bacterial metabolites induce apoptosis to kill insects
    Md. Mahi Imam Mollah, Fatema Yeasmin, Yonggyun Kim
    Journal of Asia-Pacific Entomology.2020; 23(2): 449.     CrossRef
  • Partner-specific induction of Spodoptera frugiperda immune genes in response to the entomopathogenic nematobacterial complex Steinernema carpocapsae-Xenorhabdus nematophila
    Louise Huot, Audrey Bigourdan, Sylvie Pagès, Jean-Claude Ogier, Pierre-Alain Girard, Nicolas Nègre, Bernard Duvic
    Developmental & Comparative Immunology.2020; 108: 103676.     CrossRef
  • The prophenoloxidase system in Drosophila participates in the anti-nematode immune response
    Dustin Cooper, Caitlin Wuebbolt, Christa Heryanto, Ioannis Eleftherianos
    Molecular Immunology.2019; 109: 88.     CrossRef
  • Variation in pathogenicity of different strains of Xenorhabdus nematophila; Differential immunosuppressive activities and secondary metabolite production
    Md. Ariful Hasan, Shabbir Ahmed, Md. Mahi Imam Mollah, Dongwoon Lee, Yonggyun Kim
    Journal of Invertebrate Pathology.2019; 166: 107221.     CrossRef
  • Prostaglandins and Other Eicosanoids in Insects: Biosynthesis and Biological Actions
    David Stanley, Yonggyun Kim
    Frontiers in Physiology.2019;[Epub]     CrossRef
  • Identification of entomopathogenic nematodes and symbiotic bacteria from Nam Nao National Park in Thailand and larvicidal activity of symbiotic bacteria against Aedes aegypti and Aedes albopictus
    Temsiri Yooyangket, Paramaporn Muangpat, Raxsina Polseela, Sarunporn Tandhavanant, Aunchalee Thanwisai, Apichat Vitta, Guido Favia
    PLOS ONE.2018; 13(4): e0195681.     CrossRef
  • Differential immunosuppression by inhibiting PLA2 affects virulence of Xenorhabdus hominickii and Photorhabdus temperata temperata
    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
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    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
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    Ángel M. Casanova-Torres, Upasana Shokal, Neta Morag, Ioannis Eleftherianos, Heidi Goodrich-Blair, Harold L. Drake
    Applied and Environmental Microbiology.2017;[Epub]     CrossRef
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    Simona Binda-Rossetti, Maristella Mastore, Marina Protasoni, Maurizio F. Brivio
    Journal of Invertebrate Pathology.2016; 133: 110.     CrossRef
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    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
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    Kristen E. Murfin, Amy C. Whooley, Jonathan L. Klassen, Heidi Goodrich-Blair
    BMC Genomics.2015;[Epub]     CrossRef
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    Youngjin Park
    Journal of Asia-Pacific Entomology.2015; 18(3): 459.     CrossRef
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    Seonghyeon Eom, Youngjin Park, Yonggyun Kim
    The Korean Journal of Pesticide Science.2015; 19(3): 301.     CrossRef
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    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
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NOTE] Construction and Characterisation of an Antifungal Recombinant Bacillus thuringiensis with an Expanded Host Spectrum
Qin Liu , Jong Yul Roh , Yong Wang , Jae Young Choi , Xue Ying Tao , Jae Su Kim , Yeon Ho Je
J. Microbiol. 2012;50(5):874-877.   Published online November 4, 2012
DOI: https://doi.org/10.1007/s12275-012-2201-7
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  • 6 Scopus
AbstractAbstract
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
Hyung Uk Jeong , Hye Yeon Mun , Hyung Keun Oh , Seung Bum Kim , Kwang Yeol Yang , Iksoo Kim , Hyang Burm Lee
J. Microbiol. 2010;48(4):541-545.   Published online August 20, 2010
DOI: https://doi.org/10.1007/s12275-010-0221-9
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  • 24 Scopus
AbstractAbstract PDF
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
Do Young Kim , Doo-Sang Park , Soon Bum Kwon , Moon Gyu Chung , Kyung Sook Bae , Ho-Yong Park , Young Ha Rhee
J. Microbiol. 2009;47(5):651-656.   Published online October 24, 2009
DOI: https://doi.org/10.1007/s12275-009-0109-7
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  • 18 Scopus
AbstractAbstract PDF
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
Doo-Sang Park , Hyun-Woo Oh , Sun-Yeon Heo , Woo-Jin Jeong , Dong Ha Shin , Kyung Sook Bae , Ho-Young Park
J. Microbiol. 2007;45(5):409-417.
DOI: https://doi.org/2596 [pii]
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AbstractAbstract PDF
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
Soo-Dong Woo , Jong Yul Roh , Jae Young Choi , Byung Rae Jin
J. Microbiol. 2007;45(2):133-138.
DOI: https://doi.org/2522 [pii]
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AbstractAbstract PDF
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)
Kazlm Sezen , Ismail Demir , Hatice Katl , Zihni Demirbag
J. Microbiol. 2005;43(5):463-468.
DOI: https://doi.org/2274 [pii]
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AbstractAbstract PDF
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
Hwang, Sung Hei , Yoo, Kwan Hee , Moon, Eui Sik , Cha Soung Chul , Lee, Hyung Hoan
J. Microbiol. 1998;36(4):289-295.
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AbstractAbstract PDF
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.

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