Bon-Tag koo - Academia.edu (original) (raw)

Papers by Bon-Tag koo

Research paper thumbnail of Identification of insecticidal compounds from Streptomyces sp. no. 46 = Streptomyces sp. no. 46이 생산하는 살충성 물질의 구조 동정

Journal of Korean Society of Agricul Chem Biot, 1994

Research paper thumbnail of Integration and expression of Bacillus thuringiensis crystal protein gene in chromosomal DNA of Pseudomonas strains using transposon Tn5

Korean Journal of Applied Microbiology & Biotechnology, 1991

Research paper thumbnail of Construction of Recombinant Xanthomonas campestris Strain Producing Insecticidal Protein of Bacillus thuringiensis

Journal of Microbiology and Biotechnology, 1994

An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated ... more An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated into the chromosome of a xanthan-producing bacterium, Xanthomonas campestris XP92. The cryIA(c) gene expression cassette was constructed that placed the gene between the trc promoter and rrnB transcriptional terminator. The gene was also included to prevent the expression of cryIA(c) gene in X campestris cells. Southem blot analysis confirmed the integration of the cryIA(c) gene expression cassette in chromosome of X campestris XP92 transconjugant. Expression of the insecticidal crystal protein was confirmed by Western blot analysis and bioassay against the larvae of Hyphantria cunea (Lepidoptera： Arctiidae) and Plutella xylostella (Lepidoptera：Plutellidae).

Research paper thumbnail of Identification of insecticidal compounds from Streptomyces sp. no. 46

In searching for new insecticidal compounds from soil microorganisms, strains of streptomyces spe... more In searching for new insecticidal compounds from soil microorganisms, strains of streptomyces species showed insecticidal activities on Musca domestica and Bombyx mori were selected. Compounds I-IV, which were isolated from the metabolites of no. 46 strain, were identified as piericidin , , and , respectively by UV and NMR data analyses.

Research paper thumbnail of Construction of Shuttle Promoter-probe and Expression Vectors for Escherichia coli and Bacillus subtilis, and Expression of B. thuringiensis subsp. kurstaki HD-73 Crystal Protein Gene in the Two Species

Journal of Microbiology and Biotechnology, 1991

A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the ve... more A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the vector, a useful DNA fragment, 319 bp EcoRI fragment, having strong promoter activity has been cloned from Bacillus subtills chromosomal DNA. Selection was based on chloramphenicol resistance which is dependent upon the introduction of DNA fragments allowing expression of a chloramphenicol acetyl transferase gene. The nucleotide sequence of the 319 bp fragment has been determined and the putative -35 and -10 region, ribosome binding site, and ATG initiation codon were observed. This promoter was named EB promoter and the resultant plasmid which can be used as an expression vector was named pEBP313. The crystal protein gene from B. thuringiensis subsp. kurstaki HD-73 was cloned downstream from the EB promoter without its own promoter. When the resultant plasmid, pBT313, was introduced into Escherichia coli and B. subtilis, efficient synthesis of crystal protein was observed in both cells, a...

Research paper thumbnail of Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

Journal of Microbiology and Biotechnology, 2001

To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated fro... more To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The P C27 promoter cloned from the clostridial chromosomal DNA showed a 5-fold higher promoter strength than the P SPO2 promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, P C27 exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3' end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of P C27 was distinctively repressed in the presence of glucose. Using P C27 as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolytic...

Research paper thumbnail of Cloning and expression of Serratia marcescens protease gene in Escherichia coli

Journal of Microbiology and Biotechnology, 1992

A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular se... more A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular serine protease gene was cloned in Escherichia coli. The cloned gene(pSMP18) caused specific excretion of the protease into the extracellular medium through the outer membrane of E. coli host cells. The protease purified from E. coli harboring pSMP18 was inactivated not by 100 mM EDTA but by 10 mM phenyl methyl sulfonyl flouride (PMSF). The molecular weight of the purified serine protease was about 66, 000 in the SDS-PAGE and the isoelectric point was approximately 5.7 in IEFㆍGel electrophoresis. The optimal pH and temperature for reaction of the purified serine protease were 9.5 and , respectively.

Research paper thumbnail of 국내 식물시료에서 분리한 Bacillus thuringiensis 균주의 다양성

Research paper thumbnail of Detection of cryIB genes in Bacillus thuringiensis subsp. entomocidus and subsp. subtoxicus

Journal of Microbiology and Biotechnology, 1994

To find new crystal protein genes, we screened 42 Bacillus thuringiensis strains of serovar stand... more To find new crystal protein genes, we screened 42 Bacillus thuringiensis strains of serovar standards by Southern hybridization with a cryI-specific probe which was amplified from B. thuringiensis subsp. kurstaki HDl by polymerase chain reaction (PCR). Two strains, B. thuringiensis subsp. entomocidus HD9 and subsp. subtoxicus HD109, generated weak signals under the low-stringency hybridization conditions. Further analysis with Southern hybridization revealed that the two strains contained cryIB genes which are slightly different from those of B. thuringiensis subsp. thuringiensis HD2. These results were confirmed by PCR with cryIB-specific primers followed by the restriction analysis of PCR products.

Research paper thumbnail of Distribution of cryV-type insecticidal protein genes in Bacillus thuringiensis and cloning of cryV-type genes from Bacillus thuringiensis subsp. kurstaki and Bacillus thuringiensis subsp. entomocidus

Applied and Environmental Microbiology, 1995

DNA dot blot hybridizations with a cryV-specific probe and a cryI-specific probe were performed t... more DNA dot blot hybridizations with a cryV-specific probe and a cryI-specific probe were performed to screen 24 Bacillus thuringiensis strains for their cryV-type (lepidopteran- and coleopteran-specific) and cryI-type (lepidopteran-specific) insecticidal crystal protein gene contents, respectively. The cryV-specific probe hybridized to 12 of the B. thuringiensis strains examined. Most of the cryV-positive strains also hybridized to the cryI-specific probe, indicating that the cryV genes are closely related to cryI genes. Two cryV-type genes, cryV1 and cryV465, were cloned from B. thuringiensis subsp. kurstaki HD-1 and B. thuringiensis subsp. entomocidus BP465, respectively, and their nucleotide sequences were determined. The CryV1 protein was toxic to Plutella xylostella and Bombyx mori, whereas the CryV465 protein was toxic only to Plutella xylostella.

Research paper thumbnail of Characterization of 1,925 Bacillus thuringiensis isolates from plants in Korea

Research paper thumbnail of AhlD, an N-acylhomoserine lactonase in Arthrobacter sp., and predicted homologues in other bacteria

Microbiology, 2003

Quorum sensing is a signalling mechanism that controls diverse biological functions, including vi... more Quorum sensing is a signalling mechanism that controls diverse biological functions, including virulence, via N-acylhomoserine lactone (AHL) signal molecules in Gram-negative bacteria. With the aim of isolating strains or enzymes capable of blocking quorum sensing by inactivating AHL, bacteria were screened for AHL degradation by their ability to utilize N-3-oxohexanoyl-l-homoserine lactone (OHHL) as the sole carbon source. Among four isolates, strain IBN110, identified as Arthrobacter sp., was found to grow rapidly on OHHL, and to degrade various AHLs with different lengths and acyl side-chain substitutions. Co-culture of Arthrobacter sp. IBN110 and the plant pathogen Erwinia carotovora significantly reduced both the AHL amount and pectate lyase activity in co-culture medium, suggesting the possibility of applying Arthrobacter sp. IBN110 in the control of AHL-producing pathogenic bacteria. The ahlD gene from Arthrobacter sp. IBN110 encoding the enzyme catalysing AHL degradation was...

Research paper thumbnail of Identification of Extracellular N -Acylhomoserine Lactone Acylase from a Streptomyces sp. and Its Application to Quorum Quenching

Applied and Environmental Microbiology, 2005

N -Acylhomoserine lactones (AHLs) play an important role in regulating virulence factors in patho... more N -Acylhomoserine lactones (AHLs) play an important role in regulating virulence factors in pathogenic bacteria. Recently, the enzymatic inactivation of AHLs, which can be used as antibacterial targets, has been identified in several soil bacteria. In this study, strain M664, identified as a Streptomyces sp., was found to secrete an AHL-degrading enzyme into a culture medium. The ahlM gene for AHL degradation from Streptomyces sp. strain M664 was cloned, expressed heterologously in Streptomyces lividans , and purified. The enzyme was found to be a heterodimeric protein with subunits of approximately 60 kDa and 23 kDa. A comparison of AhlM with known AHL-acylases, Ralstonia strain XJ12B AiiD and Pseudomonas aeruginosa PAO1 PvdQ, revealed 35% and 32% identities in the deduced amino acid sequences, respectively. However, AhlM was most similar to the cyclic lipopeptide acylase from Streptomyces sp. strain FERM BP-5809, exhibiting 93% identity. A mass spectrometry analysis demonstrated t...

Research paper thumbnail of Genes Encoding the N -Acyl Homoserine Lactone-Degrading Enzyme Are Widespread in Many Subspecies of Bacillus thuringiensis

Applied and Environmental Microbiology, 2002

Gram-negative bacteria can communicate with each other by N -acyl homoserine lactones (AHLs), whi... more Gram-negative bacteria can communicate with each other by N -acyl homoserine lactones (AHLs), which are quorum-sensing autoinducers. Recently, the aiiA gene (encoding an enzyme catalyzing the degradation of AHL) has been cloned from Bacillus sp. strain 240B1. During investigations in the course of the ongoing Bacillus thuringiensis subsp. morrisoni genome project, an aiiA homologue gene in the genome sequence was found. These results led to consideration of the possibility of the widespread existence of the gene in B. thuringiensis. aiiA homologue genes were found in 16 subspecies of B. thuringiensis , and their sequences were determined. Comparison of the Bacillus sp. strain 240B1 aiiA gene with the B. thuringiensis aiiA homologue genes showed high homologies of 89 to 95% and 90 to 96% in the nucleotide sequence and deduced amino acid sequence, respectively. Among the subspecies of B. thuringiensis having an aiiA gene, the subspecies aizawai , galleriae , kurstaki , kyushuensis , o...

Research paper thumbnail of Screening of Nested Deletion Mutants for DNA Sequencing by Direct Electrophoresis of Bacterial Cultures

Analytical Biochemistry, 1995

Research paper thumbnail of A Broad-Host-Range Promoter-Probe Vector, pKU20, and Its Use in Promoter Cloning and Expression of Bacillus thuringiensis Crystal Protein Gene in Pseudomonas putida

We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range ... more We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.

Research paper thumbnail of Identification of insecticidal compounds from Streptomyces sp. no. 46 = Streptomyces sp. no. 46이 생산하는 살충성 물질의 구조 동정

Journal of Korean Society of Agricul Chem Biot, 1994

Research paper thumbnail of Integration and expression of Bacillus thuringiensis crystal protein gene in chromosomal DNA of Pseudomonas strains using transposon Tn5

Korean Journal of Applied Microbiology & Biotechnology, 1991

Research paper thumbnail of Construction of Recombinant Xanthomonas campestris Strain Producing Insecticidal Protein of Bacillus thuringiensis

Journal of Microbiology and Biotechnology, 1994

An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated ... more An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated into the chromosome of a xanthan-producing bacterium, Xanthomonas campestris XP92. The cryIA(c) gene expression cassette was constructed that placed the gene between the trc promoter and rrnB transcriptional terminator. The gene was also included to prevent the expression of cryIA(c) gene in X campestris cells. Southem blot analysis confirmed the integration of the cryIA(c) gene expression cassette in chromosome of X campestris XP92 transconjugant. Expression of the insecticidal crystal protein was confirmed by Western blot analysis and bioassay against the larvae of Hyphantria cunea (Lepidoptera： Arctiidae) and Plutella xylostella (Lepidoptera：Plutellidae).

Research paper thumbnail of Identification of insecticidal compounds from Streptomyces sp. no. 46

In searching for new insecticidal compounds from soil microorganisms, strains of streptomyces spe... more In searching for new insecticidal compounds from soil microorganisms, strains of streptomyces species showed insecticidal activities on Musca domestica and Bombyx mori were selected. Compounds I-IV, which were isolated from the metabolites of no. 46 strain, were identified as piericidin , , and , respectively by UV and NMR data analyses.

Research paper thumbnail of Construction of Shuttle Promoter-probe and Expression Vectors for Escherichia coli and Bacillus subtilis, and Expression of B. thuringiensis subsp. kurstaki HD-73 Crystal Protein Gene in the Two Species

Journal of Microbiology and Biotechnology, 1991

A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the ve... more A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the vector, a useful DNA fragment, 319 bp EcoRI fragment, having strong promoter activity has been cloned from Bacillus subtills chromosomal DNA. Selection was based on chloramphenicol resistance which is dependent upon the introduction of DNA fragments allowing expression of a chloramphenicol acetyl transferase gene. The nucleotide sequence of the 319 bp fragment has been determined and the putative -35 and -10 region, ribosome binding site, and ATG initiation codon were observed. This promoter was named EB promoter and the resultant plasmid which can be used as an expression vector was named pEBP313. The crystal protein gene from B. thuringiensis subsp. kurstaki HD-73 was cloned downstream from the EB promoter without its own promoter. When the resultant plasmid, pBT313, was introduced into Escherichia coli and B. subtilis, efficient synthesis of crystal protein was observed in both cells, a...

Research paper thumbnail of Isolation of a Promoter Element that is Functional in Bacillus subtilis for Heterologous Gene Expression

Journal of Microbiology and Biotechnology, 2001

To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated fro... more To construct an efficient Bacillus subtilis expression vector, strong promoters were isolated from the chromosomal DNA libraries of Clostridium acetobutylicum ATCC 4259, Thermoactinomyces sp. E79, and Bacillus thermoglucosidasius KCTC 3400. The P C27 promoter cloned from the clostridial chromosomal DNA showed a 5-fold higher promoter strength than the P SPO2 promoter in the expression of the cat gene, and its sequence was estimated as an upstream region of the predicted hypothetical gene (tet-R family bacterial transcription regulator gene) in C. acetobutylicum. As a promoter element, P C27 exhibited putative nucleotide sequences that can bind with bacterial RNAP and the 3' end of the 16S rRNA just upstream of the start codon. In addition, the promoter activity of P C27 was distinctively repressed in the presence of glucose. Using P C27 as the promoter element, a glucose controllable B. subtilis expression vector was constructed and the lipase gene from Staphylococcus haemolytic...

Research paper thumbnail of Cloning and expression of Serratia marcescens protease gene in Escherichia coli

Journal of Microbiology and Biotechnology, 1992

A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular se... more A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular serine protease gene was cloned in Escherichia coli. The cloned gene(pSMP18) caused specific excretion of the protease into the extracellular medium through the outer membrane of E. coli host cells. The protease purified from E. coli harboring pSMP18 was inactivated not by 100 mM EDTA but by 10 mM phenyl methyl sulfonyl flouride (PMSF). The molecular weight of the purified serine protease was about 66, 000 in the SDS-PAGE and the isoelectric point was approximately 5.7 in IEFㆍGel electrophoresis. The optimal pH and temperature for reaction of the purified serine protease were 9.5 and , respectively.

Research paper thumbnail of 국내 식물시료에서 분리한 Bacillus thuringiensis 균주의 다양성

Research paper thumbnail of Detection of cryIB genes in Bacillus thuringiensis subsp. entomocidus and subsp. subtoxicus

Journal of Microbiology and Biotechnology, 1994

To find new crystal protein genes, we screened 42 Bacillus thuringiensis strains of serovar stand... more To find new crystal protein genes, we screened 42 Bacillus thuringiensis strains of serovar standards by Southern hybridization with a cryI-specific probe which was amplified from B. thuringiensis subsp. kurstaki HDl by polymerase chain reaction (PCR). Two strains, B. thuringiensis subsp. entomocidus HD9 and subsp. subtoxicus HD109, generated weak signals under the low-stringency hybridization conditions. Further analysis with Southern hybridization revealed that the two strains contained cryIB genes which are slightly different from those of B. thuringiensis subsp. thuringiensis HD2. These results were confirmed by PCR with cryIB-specific primers followed by the restriction analysis of PCR products.

Research paper thumbnail of Distribution of cryV-type insecticidal protein genes in Bacillus thuringiensis and cloning of cryV-type genes from Bacillus thuringiensis subsp. kurstaki and Bacillus thuringiensis subsp. entomocidus

Applied and Environmental Microbiology, 1995

DNA dot blot hybridizations with a cryV-specific probe and a cryI-specific probe were performed t... more DNA dot blot hybridizations with a cryV-specific probe and a cryI-specific probe were performed to screen 24 Bacillus thuringiensis strains for their cryV-type (lepidopteran- and coleopteran-specific) and cryI-type (lepidopteran-specific) insecticidal crystal protein gene contents, respectively. The cryV-specific probe hybridized to 12 of the B. thuringiensis strains examined. Most of the cryV-positive strains also hybridized to the cryI-specific probe, indicating that the cryV genes are closely related to cryI genes. Two cryV-type genes, cryV1 and cryV465, were cloned from B. thuringiensis subsp. kurstaki HD-1 and B. thuringiensis subsp. entomocidus BP465, respectively, and their nucleotide sequences were determined. The CryV1 protein was toxic to Plutella xylostella and Bombyx mori, whereas the CryV465 protein was toxic only to Plutella xylostella.

Research paper thumbnail of Characterization of 1,925 Bacillus thuringiensis isolates from plants in Korea

Research paper thumbnail of AhlD, an N-acylhomoserine lactonase in Arthrobacter sp., and predicted homologues in other bacteria

Microbiology, 2003

Quorum sensing is a signalling mechanism that controls diverse biological functions, including vi... more Quorum sensing is a signalling mechanism that controls diverse biological functions, including virulence, via N-acylhomoserine lactone (AHL) signal molecules in Gram-negative bacteria. With the aim of isolating strains or enzymes capable of blocking quorum sensing by inactivating AHL, bacteria were screened for AHL degradation by their ability to utilize N-3-oxohexanoyl-l-homoserine lactone (OHHL) as the sole carbon source. Among four isolates, strain IBN110, identified as Arthrobacter sp., was found to grow rapidly on OHHL, and to degrade various AHLs with different lengths and acyl side-chain substitutions. Co-culture of Arthrobacter sp. IBN110 and the plant pathogen Erwinia carotovora significantly reduced both the AHL amount and pectate lyase activity in co-culture medium, suggesting the possibility of applying Arthrobacter sp. IBN110 in the control of AHL-producing pathogenic bacteria. The ahlD gene from Arthrobacter sp. IBN110 encoding the enzyme catalysing AHL degradation was...

Research paper thumbnail of Identification of Extracellular N -Acylhomoserine Lactone Acylase from a Streptomyces sp. and Its Application to Quorum Quenching

Applied and Environmental Microbiology, 2005

N -Acylhomoserine lactones (AHLs) play an important role in regulating virulence factors in patho... more N -Acylhomoserine lactones (AHLs) play an important role in regulating virulence factors in pathogenic bacteria. Recently, the enzymatic inactivation of AHLs, which can be used as antibacterial targets, has been identified in several soil bacteria. In this study, strain M664, identified as a Streptomyces sp., was found to secrete an AHL-degrading enzyme into a culture medium. The ahlM gene for AHL degradation from Streptomyces sp. strain M664 was cloned, expressed heterologously in Streptomyces lividans , and purified. The enzyme was found to be a heterodimeric protein with subunits of approximately 60 kDa and 23 kDa. A comparison of AhlM with known AHL-acylases, Ralstonia strain XJ12B AiiD and Pseudomonas aeruginosa PAO1 PvdQ, revealed 35% and 32% identities in the deduced amino acid sequences, respectively. However, AhlM was most similar to the cyclic lipopeptide acylase from Streptomyces sp. strain FERM BP-5809, exhibiting 93% identity. A mass spectrometry analysis demonstrated t...

Research paper thumbnail of Genes Encoding the N -Acyl Homoserine Lactone-Degrading Enzyme Are Widespread in Many Subspecies of Bacillus thuringiensis

Applied and Environmental Microbiology, 2002

Gram-negative bacteria can communicate with each other by N -acyl homoserine lactones (AHLs), whi... more Gram-negative bacteria can communicate with each other by N -acyl homoserine lactones (AHLs), which are quorum-sensing autoinducers. Recently, the aiiA gene (encoding an enzyme catalyzing the degradation of AHL) has been cloned from Bacillus sp. strain 240B1. During investigations in the course of the ongoing Bacillus thuringiensis subsp. morrisoni genome project, an aiiA homologue gene in the genome sequence was found. These results led to consideration of the possibility of the widespread existence of the gene in B. thuringiensis. aiiA homologue genes were found in 16 subspecies of B. thuringiensis , and their sequences were determined. Comparison of the Bacillus sp. strain 240B1 aiiA gene with the B. thuringiensis aiiA homologue genes showed high homologies of 89 to 95% and 90 to 96% in the nucleotide sequence and deduced amino acid sequence, respectively. Among the subspecies of B. thuringiensis having an aiiA gene, the subspecies aizawai , galleriae , kurstaki , kyushuensis , o...

Research paper thumbnail of Screening of Nested Deletion Mutants for DNA Sequencing by Direct Electrophoresis of Bacterial Cultures

Analytical Biochemistry, 1995

Research paper thumbnail of A Broad-Host-Range Promoter-Probe Vector, pKU20, and Its Use in Promoter Cloning and Expression of Bacillus thuringiensis Crystal Protein Gene in Pseudomonas putida

We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range ... more We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.