• Journal of Industrial Technology
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• v.29 no.B
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• pp.121-125
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• 2009

New cellulase genes, named as CelV2 and CelN1, respectively, were isolated from Erwinia carotovora ATCC15713 and expressed in E. coli. The CelV2 and CelN1 gene were PCR amplified with degenerated primers and PCR products were sequenced and expressed in E. coli. Two new cellulase genes showed 97% homologies with previously reported Erwinia cellulase genes. The recombinant cellulase were purified with Ni-NTA column chromatography and its enzymatic properties were characterized. The optimum temperature of two enzymes were about $50^{\circ}C$ degree and optimum pH were around pH7.0. The newly isolated celluase genes could be used for enhancing substrate range of alcohol-producing bacteria such as Zymomonas mobilis.

• Applied Biological Chemistry
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• v.38 no.2
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• pp.111-117
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• 1995

In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E. coli MK2001, which is introduced the altered crp, is functional in the expression of lac, ara and man, in the absence of cAMP. However, the expression of mal gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mal gene expression with the altered crp gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp, cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E. coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAP5, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 bp segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.171-171
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• 2005

• Proceedings of the Zoological Society Korea Conference
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• 1995.10b
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• pp.314.1-314
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• 1995

• Proceedings of the Zoological Society Korea Conference
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• 2003.08a
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• pp.192.2-192
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• 2003

• Proceedings of the Zoological Society Korea Conference
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• 1999.10b
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• pp.289.1-289
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• 1999

• Journal of Sericultural and Entomological Science
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• v.34 no.2
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• pp.20-25
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• 1992

The location of the polyhedrin gene of Bmbyx mori nuclear polyhedrosis virus(BmNPV) was determined by using a cloned polyhedrin gene from the Autographa californica nuclear polyhedrosis virus(AcNPV) as a hybridization probe. The 7.4 Kb PstⅠ fragment DNA of Bm-NPV was cloned to plasmid pUC19 vector. A fragment containing this gene was mapped and sequenced in its entire polyhedrin reading frame. Nucleotide sequences comparison of the polyhedrin of the BmNPV to that of previously reported by Ⅰatrou(1985) revealed that the sequence varied in 10 base, Comparison of the amino acid sequence of the two structured gene revealed that coding sequence varied 74 valine to isoleucine, 76 aspargine to serine and 155 methionine to valine.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.633-639
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• 1990

The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

• Microbiology and Biotechnology Letters
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• v.38 no.2
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• pp.136-143
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• 2010

DNA fragments encoding the ketosynthase (KS) domain of polyketide synthase (PKS) genes were amplified using polymerase chain reaction (PCR) from 9 strains of Sorangium cellulosum isolated in Korea, cloned into a plasmid vector and sequenced. A total of 83 cloned DNA fragments were analyzed, and similar fragments were excluded, leaving 43 independent DNA fragments encoding the KS domains. The predicted amino acid sequences of 32 fragments were 70%-100% identical to the amino acid sequences of already known PKS genes, while the remaining 11 fragments were $\leq$67% or less identical to the known sequences, suggesting that these genes are novel PKS genes.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.126-131
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• 1994

The pchABCD genes in Pseudomonas sp. DJ-12 speciyin degradation o 4-chlorobiphenyl(4CB) were cloned in Eschericia coli. The cloned cells of E. coli CU1 and CU101 showed to produce 2,3-dihydroxybiphenyl (2,3-DHBP) from 4-chlorobiphenyl by dechlorination, as Pseudomonas so. DJ-12 produced 2,3-DHBP from both biphenyl and 4CB. In particular, E. coli CU101 transformed with the recombinant plasmid of pCU101 revealed dechlorination activity to produce 2,3-DHBP from 4CB without production of 4-chlorobenzoic acid. Therefore, the pcbAB genes (2.2 kb in size) cloned from the chromosome of Pseudomonas sp. DJ-12 were found to have dechlorination activity on 4CB to produce 2,3-DHNP.