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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
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The Microbiological Society of Korea
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Volume & Issues
Volume 38, Issue 4 - Dec 2002
Volume 38, Issue 3 - Sep 2002
Volume 38, Issue 2 - Jun 2002
Volume 38, Issue 1 - Mar 2002
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Human Cytomegalovirus Inhibition of Interferon Signal Transduction
Daniel M. Miller ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 203~203
Cytomegalovirus (CMV), a beta-herpesvirus with worldwide distribution, exhibits host persistence, a distinguishing characteristic of all herpesviruses. This persistence is dependent upon restricted gene expression in infected cells as well as the ability of productively infected cells to escape from normal cell-mediated anti-viral immunosurveillance. Type I (IFN-α/β) and type II (IFN-γ) interferons are major components of the innate defense system against viral infection. They are potent inducers of MHC class I and II antigens and of antigen processing proteins. Additionally, IFNS mediate direct antiviral effects through induction effector molecules that block viral infection and replications such as 2′, 5-oligoadenylate synthetase (2, 5-OAS). IFNS function through activation of well-defined signal transduction pathways that involve phosphorylation of constituent proteins and ultimate formation of active transcription factors. Recent studies have shown that a number of diverse viruses, including CMV, EBV, HPV mumps and Ebola, are capable of inhibiting IFN-mediated signal transduction through a variety of mechanisms. As an example, CMV infection inhibits the ability of infected cells Is transcribe HLA class I and II antigens as well as the antiviral effector molecules 2, 5-OAS and MxA I. EMSA studies have shown that IFN-α and IFN-γ are unable to induce complete signal transduction in the presence of CMV infection, phenomena that are associated with specific decreases in JAKl and p48. Viral inhibition of IFN signal transduction represents a new mechanistic paradigm for increased viral survival, a paradigm predicting widespread consequences in the case of signal transduction factors common to multiple cytokine pathways.
Enzyme Activities Related to the Methanol Oxidation of Mycobacterium sp. strain JCl DSM 3803
Youngtae Ro ; 김응빈 ; 김영민 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 209~209
Mycobacterium sp. strain JCl DSM 3803 grown in methanol showed no methanol dehydrogenase or oxidase activities found in mast methylotrophic bacteria and yeasts, respectively. Even though the methanol-grown cells exhibited a little methanol-dependent oxidation by cytochrome c-dependent methanol dehydrogenase and alcohol dehydrogenase, they were not the key enzymes responsible for the methanol oxidation of the cells, in that the cells contained no c-type cytochrome and the methanol oxidizing activity from the partially purified alcohol dehydrogenase was too low, respectively. In substrate switching experiments, we found that only a catalase-peroxidase among the three types of catalase found in glucose-grown cells was highly expressed, in the methanol-grown cells and that its activity was relatively high during the exponential growth phase in Mycobacterium sp. JCl. Therefore, we propose that catalase-peroxidase is an essential enzyme responsible for the methanol metabolism directly Of indirectly in Mycobacterium sp. JCl.
Intracellular Posttranslational Modification of Aspartyl Proteinase of Candida albicans and the Role of the Glycan Region of the Enzyme
나병국 ; 송철용 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 218~218
Using two drugs, tunicamycin and brefeldin A, which affect protein processing, we investigated the intracellular processing mechanism of secreted aspartyl proteinase 1 (SAPl) of Candide albicans. Three intracellular forms of SAPI were detected by immunoblotting using menoclonal antibody (MAb) CAPl. Their molecular weights were approximately 40, 41 and 45 kDa, respectively. The 41 kDa protein is a glycoprotein and may be the same as the extracellular form judging by its molecular mass. The 40 kDa protein was the unglycosylated form and its molecular mass coincided with deglycosylated SAPl and the 45 kDa protein was also the unglycosylated form. Neither the 40 and 45 kDa proteins were detected in the culture supernatant of C. albicans. These suggested that the 40 and 45 kDa proteins might be intracellular precursor forms of SAPI. These results show that SAPI is translated as a 45 kDa precusor form in the endoplasmic reticulum and the 45 kDa precursor farm undergoes proteolytic cleavage after translocation into the Golgi apparatus, generating the 40 kDa precursor form. This 40 kDa precursor is converted into a 41 kDa mature form through glycosylation in the Golgi apparatus. The mature form of the 41 kDa protein is sorted into secretary vesicles and finally released into the extracellular space through membrane fusion. When the glycan region of SAPl was digested with N-glycosidase F, both stability and activity of the enzyme decreased. These results indicate that the glycan attached to the enzyme may, at least in parti be related to enzyme stability and activity.
Purification and Characterization of Chitinase from a Marine Bacterium, Vibrio sp. 98CJ11027
박신혜 ; 이정현 ; 이홍금 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 224~224
Chitin-degrading marine bacterial strain 98CJ11027 was isolated from bryozoa from the coastal area of Cheju Island, Korea, and identified as a member of the genus Vibrio. The molecular mass of the main extracellular chitinase (chitinase I), purified from strain 98CJ11027, was estimated to be 98 kDa. The optimal condition for chitinase I activity is pH 6.0 and 45℃. The activity was inhibited by
. Chitinase I displayed the hydrolysis type of chitobiosidase and catalyzed reversed hydrolysis leading to the synthesis of tetraacetylchitotetraose.
Genomic Organization of Penicillium chrysogenum chs4, a Class Ⅲ Chitin Synthase Gene
박윤동 ; 이명숙 ; 남경준 ; 박범찬 ; 배경숙 ; 박희문 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 230~230
Class Ⅲ chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class Ⅲ chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5′flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.
Isolation and Characterization of Aeromons hydrophila PBl6 and Properties of Synthetic Wastewater Degradation
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 235~240
Protease producing bacterium, PB16 was isolated from food processing wastewater sludge and paddy field soil samples and selected by the clear zone and enzyme activity test. The isolate was gram negative, rod type and its protease productivity was 6.49 U/ml. As a result of API20NE kit test and 16S rDNA sequencying, the isolated PB16 was identified as Aeromonas hydrophila (99%). The growth rate (
) was 0.21 in synthetic waste water only and 0.26 in synthetic waste water containing vitamin and mineral using a bioscreen C. Synthetic wastewater removal rate was 59 and 87%, respectively after 1 and 3 day reaction (intial CODcr was 2,472 mg/l).
Analysis of the Dual Promoters and the H₂O₂-responsive Element of the catA Gene Encoding Catalase A in Streptomyces coelicolor
Jo, Yu Hui ; Han, Ji Suk ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 239~239
Purification and Characterization of an Endo-
-1,3-1,4-Glucanase from Escherichia coli(pLL200K)
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 241~246
A gene coding for endo-
-1,3-1,4-glucanase of Bacillus circulans was subcloned into Escherichia coli Ml5 using pQE30 as an expression vector. Endo-
-1,3-1,4-glucanase produced by the recombinant expression plas-mid pLQ43 was intactly purified to a single protein through a nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography method. The molecular mass of the purified enzyme was estimated to be 28 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme activity were pH 6.8 and
, respectively. This enzyme was fairly stable in the pH ranging 5.5~7.5 and at the temperatures lower than
. The enzyme appeared to be sensitive to most of the metal ions, especially to
, and also to methanol, ethanol, isopropanol or 1-butanol at a concentration of 10%(v/v).
Roles of the meta- and the ortho-Cleavage Pathways for the Efficient Utilization of Aromatic Hydrocarbons by Sphingomonas yanoikuyae Bl
송정민 ; 김영민 ; Gerben J. Zylstra ; 김응빈 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 245~245
Catabolic pathways for the degradation of various aromatics by Sphingomonas yanoikuyae Bl are intertwined, joining at the level of substituted benzoates, which are further degraded vita ring cleavage reactions. The mutant strain EK497, which was constructed by deleting a large DNA region containing most of the genes for biphenyl, naphthalene, m-xylene, and m-toluate degradation, was unable to grow on all of the aromatics tested except for benzoate as the sole source of carbon and energy.S. yanoikuyae EK497 was found to possess only catechol ortho-ring cleavage activity due to deletion of the genes for the meta-cleavage pathway. Wild-type S. yanoikuyae Bl grown on benzoate has both catechol orthoand meta-cleavage activity. However, m-xylene and m-toluate, which are metabolized through methylbenzoate, and biphenyl, which is metabolized through benzoate, induce only the meta-cleavage pathway, suggesting the presence of a substrate-dependent induction mechanism.
Cellular Responses of the TNT-degrading Bacterium, Stenotrophomonas sp. OK-5 to Explosive 2,4,6-Trinitrotoluene (TNT)
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 247~253
The cellular responses of TNT-degrading bacterium, Stenotrophomonas sp. OK-5 to explosive 2,4,6-trini-trotoluene (TNT) as an environmental contaminant were examined. Survival of the strain OK-5 with time in the presence of different concentrations of TNT under sublethal conditions was monitored, and viable counts paralleled the production of the stress shock proteins in this bacterium. Total cellular fatty acids analysis showed that strain OK-5 produced or disappeared several different kinds of lipids when grown on TNT media than when grown on TSA. Under scanning electron microscope, the cells treated with 0.5 mM TNT for 12 hrs showed irregular rod shapes with wrinkled surfaces. Analyses of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that several stress shock proteins including 70 kDa DnaK and 60 kDa GroEL in strain OK-5 were newly synthesized at different TNT concentrations in exponentially growing cultures. 2-D PAGE of soluble protein fractions from the culture of OK-5 exposed to TNT demonstrated that approximately 300 spots were observed on the silver stained gel ranging from pH 3 to pH 10. Among them, 10 spots significantly induced and expressed in response to TNT were selected and analyzed. As the result of internal amino acid sequencing with ESI-Q TOF, two proteins, spot #1 and spot #10 were assigned the DnaK protein XF2340 of Xylella fastidiosa and stress-induced protein of Mesorhizobium loti, respectively.
Simultaneous Utilization of Two Different Pathways in Degradation of 2,4,6-Trinitrotoluene by White Rot Fungus Irpex lacteus
김현영 ; 송홍규 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 250~250
This study confirmed that white rot fungus Irpex lacteus was able to metabolize 2,4,6-trinitrotoluene (TNT) with two different initial transformations. In one metabolic pathway of TNT a nitro group was removed from the aromatic ring of TNT. Hydride-Meisenheimer complexes of TNT (H/sup -/-TNT), colored dark redo were confirmed as the intermediate in this transformation by comparison with the synthetic compounds. 2,4-Dinitrotoluene as a following metabolic product was detected, and nitrite produced by denitration of
-TNT supported this transformation. In the other TNT pathway, nitro groups in TNT were successively reduced to amino groups via hydroxylamines. Hydroxylamino-dinitrotoluenes and amino-dinitrotoluenes were identified as the intermediates. The activity of a membrane-associated aromatic nitroreductase was detected in the cell-free extract of I. lacteus. This enzyme catalyzed the nitro group reduction of TNT with NADPH as a cofactor, Enzyme activity was not observed in the presence of molecular oxygen.
Purification and Characterization of Extracellular Protease form Psychrotrophic Antarctic Bacteria
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 254~259
A psychrotrophic bacterium was isolated from Antarctic marine sediment and identified as Shewanella sp. species based on the biochemical properties and 16S rRNA sequence, and designated as Shewanella sp. L93. Extracellular protease produced by this strain was purified through ammonium sulfate precipitation, High-Q column chromatography, first gel permeation chromatography, BioScale Q2 ion exchange chromatography and second gel permeation chromatography, and basic properties of this enzyme were investigated.
Measurement of Antiviral Activities Using Recombinant Human Cytomegalovirus
송병학 ; 이규철 ; 이찬희 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 255~255
For rapid and sensitive measurement of antiviral activities, application of a recombinant virus containing firefly luciferase gene was attempted. Recombinant human cytomegalovirus (HCMV) containing luciferase gene driven by HCMV late gene pp28 promoter (HCMV/pp28-luc) was used to test the antiviral activities of three known compounds and the result was compared with results from the conventional plaque assay for measuring the production of infectious viruses. When human fibroblast cells were infected with HCMV/pp28-luc, luciferase activity was observed at 2 days after infection and reached maximum at 6 days after infection, whereas the production of infectious virus was maximal at 4 days after infection. The antiviral activities of ganciclovir, acyclovir, and papaverine were measured in HFF cells infected with HCMV/PP28-luc and the luciferase activity was compared with the infectious virus titers. Luciferase activity decreased as the concentration of ganciclovir or papaverine increased, while there was a slight decrease in luciferase activity with acyclovir. The level of the decrease in Luciferase activity was comparable to the level of decrease in the production of infectious virus. Therefore, the antiviral assay using recombinant virus HCMV/pp28-luc resulted in sensitivity similar to the conventional plaque assay with a significant reduction in assay time.
Subcloning and DNA Sequencing of the Phenol Regulatory Genes in Ralstonia eutropha JMP134
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 260~266
In this study, chromosomal DNA fragment related to the regulation of phenol metabolism in Ralstonia eutropha JMP 134 was cloned and sequenced. The result has shown that two open reading frames (ORF1 and ORF2) exist on this regulatory region. ORF1, which initiates from 454 bp downstream of the stop codon of the phenol hydroxylase genes, was found to be composed of 501 amino acids. ORF2, whose start codon is overlapped with the stop codon of ORFl, was found to contain 232 amino acids. The comparison of amino acid sequences with other proteins has revealed that ORF1 belongs to the family of NtrC transcriptional activator, whereas ORF2 shares high homology with the family of GntR protein, which is known to be a negative regulator. ORF1 and ORF2 were designated as a putative positive regulator, phlR2 and a negative regulator phlA, respectively. Possible regulatory mechanisms of phenol metabolism in this strain was discussed.
Iron Increases Susceptibilities of Pseudomonas aeruginosa to Ofloxacin by Increasing the Permeability
김숙영 ; 김진숙 ; 남혜란 ; 정유선 ; 이연희 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 265~265
Iron increased the susceptibilities of clinical isolates Pseudomonas aeruginosa to quinolones. In the presence of iron, increased susceptibilities to ofloxacin were observed in twenty-six out of thirty isolates and with no change in four isolates. In the case of norfloxacin, iran increased susceptibilities of twelve isolates but did not render any change in eighteen isolates. In the case of ciprofloxacin, iron decreased the MICs (Minimal Inhibitory Concentration) of twenty isolates, increased the MIC of one isolate, and did net change the MICs of nine isolates. To find out how iron increased susceptibility to ofloxacin, bacterial cells were grown in Muller Hinton (MH) media and succinate minimal media (SMM) to induce iran acquisition systems and the intracellular ofloxacin concentrations were assayed in the presence of iron. The addition of iron to the media decreased the MICs of cells whether they were grown in MH or SMM. Siderophores, carbonyl cyanide m-chlorophenylhydrazone (an inhibiter of proton motive force), and ouabain (an inhibitor of ATPase) did not decrease the effect of iron. Results suggested that the increase in the intracellular ofloxacin concentration by iron is accomplished not by decreasing the efflux but by increasing the of ofloxacin permeability.
A Genomics Tool for Microbial Genome Comparison Using BLAST/FASTA
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 267~275
We have developed GComp as an analysis tool for microbial genome comparison. This tool exploits BLAST or FASTA as a preprocessing program for local alignments to detect homologous regions, parses the homology search results, and generates tables and files to show homology relationship between two genomes at a glance. The interface for graphical representation of the comparative genomic analysis has been also implemented. Our test cases shows that the program can be useful in practice for intuitive and quantitative comparison of microbial genome sequence pairs as well as self-genome analysis. A few additional features have been devised and designed, which will be added in the further development.
Penetration of HEp-2 and Chinese Hamster Ovary Epithelial Cells by Escherichia coli Harbouring the Invasion-Conferring Genomic Region from Salmonella typhimurium
박정욱 ; 황상구 ; 문자영 ; 조용권 ; 김동완 ; 정용기 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 270~270
Pathogenic Salmonella typhimurium can invade the intestinal epithelium and cause a wide range of diseases including gastroenteritis and bacteremia in human and animals. To identify the genes involved in the infection, the invasion determinant was obtained from S. typhimurium 82/6915 and was subcloned into pGEM-7Z. A subclone DHl (pSV6235) invaded HEp-2 and Chinese hamster ovary epithelial cells and contained a 4.4 kb fragment of S. typhimurium genomic region. Compared with the host strain E. coli DHl, the subclone DHl (pSV6235) invaded cultured HEp-2 and Chinese hamster ovary cells at least 75- and 68-fold higher, respectively. The invasion rate of E. coli DHl for the cells significantly increased by harbouring the genomic region derived from pathogenic S. typhimurium 82/6915.
Cloning and Sequence Analysis of the xyIL Gene Responsible for 4CBA-Dihydrodiol Dehydrogenase from Pseudomonas sp. S-47
박동우 ; 이상만 ; 가종옥 ; 김지경 ;
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 275~275
Pseudomonas sp. S-47 is capable of catabolizing 4-chlorobenzoate (4CBA) as carbon and energy sources under aerobic conditions via the mesa-cleavage pathway. 4CBA-dioxygenase and 4CBA-dihydrodiol dehydrogenase (4CBA-DD) catalyzed the degradation af 4CBA to produce 4-chlorocatechol in the pathway. In this study, the xylL gene encoding 4CBA-DD was cloned from the chromosomal DNA of Pseudomonas sp. S-47 and its nucleotide sequence was analyzed. The xylL gene was found to be composed of 777 nucleotide pairs and to encode a polypeptide of 28 kDa with 258 amino acid residues. The deduced amino acid sequence of the dehydrogenase (XylL) from strain S-47 exhibited 98% and 60% homologies with these of the corresponding enzymes, Pseudomonas putida mt-2 (XyIL) and Acinetobacter calcoaceticus (BenD), respectively. However, the amino arid sequences show 30% or less homology with those of Pseudomonas putida (BnzE), Pseudomonas putida Fl (TodD), Pseudomonas pseudoalcaligenes KF707 (BphB), and Pseudomonas sp. C18 (NahB). Therefore, the 4CBA-dihydrodiol dehdrogenase of strain S-47 belongs to the group I dehydrogenase involved in the degradation of mono-aryls with a carboxyl group.
Subgrouping of Pseudomonas aeruginosa Isolates by Resemblance Coefficiency of the Cellular Fatty Acid Analysis
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 276~280
To find the applicability of resemblance of the fatty acid methy1 ester (FAME) profile analysis to the primary examination for the tracing P. aeruginosa contamination in the environment and water plants, we designed this study. Of6l P. aeruginosa isolates including 3 reference strains, 4 fecal or soil isolates and 54 water isolates, the resemblance coefficiencies of the FAMEs were calculated as euclidian distance (ED) and expressed as dendrogram. The 61 strains were clustered as the same group in the ED 8.4, and divided into 4 subgroups in the ED 6.9. The water isolates from Han River were classified into distinctively different subgroups from reference strains. These results provide the possibility of the dendrogram from FAME profile as a useful tools for tracing contamination as well as for identifying the species, especially in case of P. aeruginosa.
Detection of Vibrio vulnificus in Fish Farm and Bactericidal Methods on this Bacteria
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 281~286
Detection of Vibrio vulnificus in fish farm and searching for the bactericidal methods on this bacteria were studied. To detect this microorganism in sea water, mud, fish and mussels, selective isolation methods and detection of vvhA gene were used from January to October,2000. V. vulnificus was detected from May when the water temperature was over
. From June to September, higher than
, this bacteria could be isolated from most of the samples. Freezing and refrigerating did not inhibit the growth of V. vulnificus. Citric acid did not show the bactericidal effect, but more than 500 mg/l of EDTA did. With the aid of UV and photocatalyst,
showed bactericidal effect after 15 minute treatment. Photocatalytic system consisted of glass bead coated with
and UV illumination showed bactericidal effect on V. vulnificus at the turnover rate of 0.2/min.
The Bacterial Community Structure in Cheonho Reservoir Dominated by Cyanobacteria
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 287~292
The composition of bacterial communities was detected in surface water of Cheonho Reservoir dominated by cyanobacteria, using fluorescent in situ hybridization (FISH) method. Total bacterial numbers were very high ranging from 0.6～
, whereas the ratio of Eubacteria to total bacteria was 29.8~45.8%, which was lower than that in other freshwater ecosystems. On average only 2.1% of DAPI-stained bacteria were detected by FISH with probes for
-groups, respectively. Unknown eubacteria which was not bound to any probes except EUB 338, was relatively high. On the other hand, the Cytophaga-Flavobacterium group increased following the change of dominant species from Anabaena sp. to Microcystis sp. This result showed that bacterial communities could be affected by phytoplanktons, especially cyanobacteria.
Phylogenetic Analysis of Oligotrophic Bacteria Found in Potable Groundwater
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 293~298
In order to investigate the ecological aspect of bacteria on groundwater, water samples were collected from various regions. Total of 318 strains were isolated from diluted nutrient broth (DNB) agar medium, and investigated their growth pattern on nutrient broth (NB) medium. As a result, all the isolated strains were divided into two groups, NB and DNB organisms. Growth of DNB organisms were suppressed in full strength NB medium but not in DNB medium, which were called oligotrophic bacteria in this study. Proportion of DNB organisms occurred in the frequency of 50-98% in potable groundwaters (CW, CJ, DPG, CJG1), however, it was 23,46% in polluted site (TJ, NPG1). One hundred and two strains were identified as oligotrophic bacteria and their phylogenetic characteristics were determined by using 16S rDNA sequencing. Based on the phylogenetic analysis, they were found to fall into three major phylogenetic groups: belonging to the Proteobacteria
-(3 strains) subdivisions. The phylogenetic analysis suggested that microbial diversity of potable groundwater is more complex than that obtained in the past investigation.
Organization of Antibiotic Resistance Gene Cluster of Multi-Drug Plasmid in Clinically Isolated Salmonella Enteritidis Strain
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 299~305
Clinically isolated Salmonella Enteritidis strain has a multi drug resistance plasmid, which confers ampicillin, chloramphe-nicol, sulfonamide, streptomycin and tetracycline, named pCAST2. We cloned a 7 kb Sacl fragment of pCAST2 which has sulfonamide, streptomycin and tetracycline resistance genes. The 7 kb SacI fragment showed the organization of sulII-strA-strB-tetR-tetA gene cluster which is different from the other clusters reported previously. In this study, we presented the method to detect this cluster by PCR analysis and showed that this cluster was found in Salmonella strains occurred sporadically at Kyungpook province in 2002.
A Rapid Detection of Methicillin-Resistant Staphylococci by Polymerase Chain Reaction
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 306~311
PCR of the mecA gene for the rapid detection of methicillin-resistant staphylococci was perfomed and compared with the antibiotic sensitivity test. A total of 43 strains of staphylococi from clinical specimens were used in this study. An antibiotic sensitivity test by the agar dilution method of NCCLS (The National Commitee for Clinical Laboratory Standard) was performed for the strains. Among them, 39 isolates were methicillin-resistant (MRS), and 4 isolates were methicillin-susceptible (MSS). With the exception for one strain (Staphylococcus cohnii, HRC2-4), all MRS strains amplified the expected 533 bp fragments of the mecA gene by PCR, However, one strain (Staphylococcus aureus, HSA1-10) that was classified as a sensitive strain by the antibiotic sensitivity test was mecA positive by PCR. All 35 methicillin-resistant Staphylococcus aureus (MRSA) strains were mecA positive, but overall, concordance between the results of the mecA PCR and antibiotic sensitivity test was 95.6%.
Optimization of Cell Culture Condition for Erythritol Production by Penicillium sp. KJ8l
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 312~317
Erythritol is of interest as a low calorie sweetner. Penicillium sp. KJ8l was screened for erythritol producer in nature. The effect of culture conditions on erythritol production by Penicillium sp. KJ81 was examined. This strain produced about 12 g/l erythritol and a small amout of glycerol. Erythritol was not produced from mannitol, arabinose, sorbitol, and xylose but from glucose, sucrose, fructose, mannose, lactose, maltose, and galactose. This strain was able to produce erythritol in a medium containing 60% sucrose but demonstrated the highest productivity of erythritol in a 30% sucrose medium. The highest yield in Penicillium sp. KJ8l was obtained when 0.5% ammonium sulfate was added to the medium containing 30% sucrose and 0.5% yeast extract. Penicillium sp. KJ81 produced 28.2 g/l erythritol when this strain was cultured in the medium containing 30% sucrose, 0.5% yeast extract, 0.5%
under the condition of 1 vvm aeration and 200 rpm agitation at
for 10 days in 5ι jar fermentor.
Construction of the Phosphate-Limitation Inducible Expression Vector Containing the phoA Promoter of Enterobacter aerogenes
The Korean Journal of Microbiology, volume 38, issue 4, 2002, Pages 318~321
To induce recombinant protein under phosphate restricted conditions such as soil, we have constructed the expression vector (pEAAP) with phoA gene promoter of Enterobacter aerogenes. To construct the pEAAP, deletion of the T7 promoter and lac operator from pET-22b(+) by BglII-XhoI digestion and addition of the phoA gene promoter (containing the pho box) were performed. To test pEAAP as an expression vector controled by phosphate limitation, pEAPHY1 was constructed with the phytate gene (Bsa-phy1) of Bacillus subtillis var. amyloliquefaciens (KCTC 8913P). Under the phosphate-limitation condition, CK-PHY1 ( Escherichia coli JM109 was transformed with pEAPHY1) expressed the 41 kD Bsa-Phy1 . Also CK-PHY1 formed the clear zone in solid medium containing phytate as a sole phosphate source.