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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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Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 44, Issue 4 - Dec 2008
Volume 44, Issue 3 - Sep 2008
Volume 44, Issue 2 - Jun 2008
Volume 44, Issue 1 - Mar 2008
Selecting the target year
Identification and Characterization of External Copper Responsive Genes of Deinococcus radiodurans
Joe, Min-Ho ; Lim, Sang-Yong ; Jung, Sun-Wook ; Song, Du-Sub ; Choi, Young-Ji ; Kim, Dong-Ho ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 169~177
Global gene expression of Deinococcus radiodurans, a highly radiation resistant bacterium, in response to excess copper was analyzed by using oligonucleotide microarray chip. Among 3,187 open reading frames of D. radiodurans, seventy genes showed a statistically significant expression ratio of at least 2-fold changes under growth conditions of excess copper; 64 genes were induced and 6 genes were reduced. Especially, two operons (
) presumably involved in the iron transport and utilization were the most highly induced genes by excess copper. A quantitative real-time PCR assay revealed that DRB00l4 and DRB0125 are highly transcribed responding to excess copper and 2,2'-dipyridyl, an iron chelator. In addition, the transcription of both genes was not changed by excess iron and bathocuproine disulphonate, a copper chelator. These results suggested that the copper metabolism may be closely connected with the iron transport and utilization in D. radiodurans. However, the disruption of each gene, DRB00l4 and DRB0125, did not affect the copper and radiation resistance, the most well-known character of this organism.
Expression, Purification and Antiserum Production of the Avian Influenza H9N2 Virus HA and NA Proteins
Lee, Hyun-Ji ; Song, Byung-Hak ; Kim, Jeong-Min ; Yun, Sang-Im ; Kim, Jin-Kyoung ; Kang, Young-Sik ; Koo, Yong-Bum ; Jeon, Ik-Soo ; Byun, Sung-June ; Lee, Youn-Jeong ; Kwon, Jun-Hun ; Park, Jong-Hyeon ; Joo, Yi-Seok ; Lee, Young-Min ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 178~185
Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.
Development of Trans-Splicing Aptazyme Which Can Specifically Modify Hepatitis C Virus Genome
Kim, Ju-Hyun ; Lee, Chang-Ho ; Jang, Sun-Young ; Lee, Seong-Wook ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 186~192
For the development of specific and effective basic genetic materials to inhibit replication of hepatitis C virus (HCV), HCV genome-targeting trans-splicing aptazyme, which activity is allosterically regulated by a specific ligand, was developed. The aptazyme was designed to be comprised of sequence of RNA aptamer to the ligand, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding the ligand to the aptamer, and trans-splicing ribozyme targeting +199 nt of HCV IRES. Especially, when the aptamer and the communication module was inserted at both P6 and P8 catalytic domain of the specific ribozyme, allosteric activity of the aptazyme was the most induced. The aptazyme was shown to induce activity of trans-splicing reaction specifically and efficiently only in the presence of the specific ligand, but neither in the absence of any ligand nor in the presence of control ligand. This aptazyme can be used as a specific and effective genetic agent against HCV, and a tool for the isolation of anti-HCV lead compounds.
Functional Role of
in 23S rRNA Methylation, Which is in N-Terminal End Region of ErmSF
Jin, Hyung-Jong ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 193~198
ErmSF is one of the proteins which are produced by Streptomyces fradiae to avoid suicide by its autogenous macrolide antibiotic, tylosin and one of ERM proteins which are responsible for transferring the methyl group to
(Escherichia coli coordinate) in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confers the antibiotic resistance on microorganisms ranging from antibiotic producers to pathogens. ErmSF contains an extra N-terminal end region (NTER), which is unique to ErmSF and 25% of amino acids of which is arginine known well to interact with RNA. Noticeably, arginine is concentrated in
and functional role of each arginine in this motif was investigated through deletion and site-directed mutagenesis and the activity of mutant proteins in cell R60 and R61 was found to play an important role in enzyme activity through the study with deletion mutant up to R60 and R61. With the site-directed mutagenesis using deletion mutant of 1 to 59 (R60A, R61A, and RR60, 61AA), R60 was found more important than R61 but R61 was necessary for the proper activity of R60 and vice versa. And these amino acids were presumed to assume a secondary structure of
Degradation of Bisphenol A and Removal of Its Estrogenic Activity by Two Laccase Transformants of Irpex lacteus
Kim, Yun-Jung ; Song, Hong-Gyu ; Choi, Hyoung-T. ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 199~202
A white rot fungus Irpex lacteus produced lignin degrading enzymes, which showed degrading activity against various recalcitrant compounds. However, laccase, one of the lignin degrading enzymes, was too low to be assayed by spectrophotometry using o-tolidine as the chromogenic substrate in this fungus under various culture conditions. A laccase expression vector was constructed using a cDNA from Phlebia tremellosa with the constitutively expressed promoter of glyceraldehydes-3-phosphate dehydrogenase gene, and introduced into I. lacteus by the restriction enzyme mediated integration transformation through the protoplast-
procedure. Two transformants showed highly increased laccase activities at the early growth phase in the minimal liquid medium, and they not only degraded bisphenol A, a notorious endocrine disrupting chemical, but also removed the estrogenic activity effectively.
Rapid Detection for Shiga Toxin Type 1 (Stxl) by Using Two-Step Ultra-Rapid Real-Time (URRT) PCR
Kim, Il-Wook ; Kang, Min-Hee ; Kwon, Soon-Hwan ; Cho, Seung-Hak ; Yoon, Byoung-Su ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 203~211
Rapid detection-method for Shiga toxin type 1 that was produced from Shiga toxin-producing Escherichia coli (STEC) was developed by two-step ultra-rapid real-time (URRT) PCR. The specific primers were deduced from 80 bp stable region of stx type 1 (stxl) gene among various informations of STEC strains. URRT PCR is a microchip-based real-time PCR using 6
of reaction volume with extremely short denaturation step and annealing/extension step (1 sec, 3 sec, respectively) in each cycle of PCR. Using the stx1-specific URRT PCR, 35 cycled PCR were finished in time of 6 min and 38 see, also measured 7 min and 28 see including melting temperature (Tm) analysis. The detection-limit of stxl-specific URRT-PCR was estimated until 3 colony forming units / PCR with products with stable Tm at
. In the applications to various STEC strains and contaminated genomic DNAs, stx1-specific URRT-PCR were tested and shown that it would be expected an useful method for the rapid detection of stx1-coded STEC strains.
Development of Prevotella nigrescens ATCC
-Specific PCR Primers
Song, Soo-Keun ; Yoo, So-Young ; Kim, Mi-Kwang ; Kim, Hwa-Sook ; Lim, Sun-A ; Kim, Do-Kyung ; Park, Jae-Yoon ; Kook, Joong-Ki ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 212~220
A Pn10 DNA probe was introduced as a Prevotella nigrescens ATCC
-specific DNA probe. In that study, the specificity of the Pn10 was tested with only type or reference strains of 5 oral bacterial species. The purpose of this study is to evaluate the specificity of the Pn10 using the wild type strains of P. nigrescens and is to develop the P. nigrescens ATCC
-specific PCR primers based on the nucleotide sequence of the Pn10. The specificity of the Pn10 DNA probe was determined by Southern blot analysis. The nucleotide sequence of Pn10 DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pn10 DNA probe were hybridized with the genomic DNAs from P. nigrescens ATCC
and KB6. The Pn10 homologous region, KB6-Pn10, of P. nigrescens KB6 was cloned by PCR and sequenced. The Pn10 and KB6-Pn10 DNA fragments were consisted of 1,875 bp and 1,873 bp, respectively. The percent identity of the two was 98.8% and the divergence of them was 0.6%. The two primer sets (Pn10-F-AC/ Pn10-R-AC and Pn10-F-A/ Pn10-R-A), designed base on the nucleotide sequences of Pn10 DNA probe, were specific to the P. nigrescens ATCC
. The two PCR primer sets could detect as little as 4 pg of genomic DNA of P. nigrescens ATCC
. These results indicate that the two PCR primer sets have proven useful for the identification of P. nigrescens ATCC
, especially with regard to the maintenance of the strain.
Serotype Variations of Agglutinogen and Fimbriae in the Korean Isolates of Bordetella pertussis
Jung, Sang-Oun ; Moon, Yu-Mi ; Sung, Hwa-Young ; Kang, Yeon-Ho ; Yu, Jae-Yon ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 221~227
Bordetella pertussis is pathogenic bacteria causing pertussis, a infectious respiratory disease for the infants. The incidence rate of pertussis was significantly decreased after introduction of vaccine. However, increased pertussis cases are recently reported in several countries with high vaccine coverage. One of the inferred reasons is genotype or serotype variation between circulating strains and vaccine strains. Therefore, it is required to confirm the variation status of the isolates by genotype or serotype analysis and the possibility of pertussis outbreak in Korea should be estimated. For this, the serotype variations of the isolates from
were investigated in agglutinogen and fimbriae. As the result, the most frequent serotype in the isolated strains was agglutinogen 1 and fimbriae 2 serotypes. Moreover, serotype transition from vaccine serotypes to non-vaccine serotypes was observed. Especially, the transition pattern of agglutinogen serotype was directed to increase a different type (agg 1) from the vaccine type (agg 1,2). However, in case of fimbriae, the same type (fim 2) with vaccine strain was increased. These results were also observed in other countries with increasing incidence of pertussis. For more predictable results to know increasing possibility of pertussis incidence in Korea, the studies on genetic variations of antigenic determinant genes and prevalence of antibody titer in normal population should be performed in the further.
Real-Time RT-PCR for Validation of Reovirus Type 3 Safety During the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals
Lee, Dong-Hyuck ; Jeong, Hyo-Sun ; Kim, Tae-Eun ; Oh, Seon-Hwan ; Lee, Jung-Suk ; Kim, In-Seop ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 228~236
Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to Reovirus type 3 (Reo-3), and there are several reports of Reo-3 contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the Reo-3 safety, a real-time RT-PCR method was developed for quantitative detection of Reo-3 in cell lines, raw materials, manufacturing processes, and final products as well as Reo-3 clearance validation. Specific primers for amplification of Reo-3 RNA was selected, and Reo-3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be
. The real-time RT-PCR method was proven to be reproducible and very specific to Reo-3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with Reo-3. Reo-3 RNA could be quantified in CHO cell as well as culture supernatant. When the real-time RT-PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of Reo-3.
Comparison of Phylogenetic Characteristics of Bacterial Populations in a Quercus and Pine Humus Forest Soil
Han, Song-Ih ; Cho, Min-Hye ; Whang, Kyung-Sook ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 237~243
Chemical and microbial characteristics of bacterial populations were investigated in a quercus and pine humus forest soil. Soil pH was
from each sample of a quercus and pine humus forest soil; C/N ratio of humus forest soil was
, respectively. Total organic acid was investigated as 69.57 mM/g dry soil and 53.72 mM/g dry soil in each humus forest soil. Glutamine, pyruvate, succinate, lactic acid and acetic acid of pine humus forest soil were
times higher than those of quercus humus forest soil. As we evaluated phylogenetic characteristics of bacterial populations by 16S rRNA-ARDRA analysis with DNA extracted from each humus forest soil. Based on the 16S rRNA sequences, 44 clone from ARDRA groups of quercus humus forest soil were classified into 7 phyla:
-Proteobacteria, Acidobacteria, Actinobacteria, and Firmicutes. Thirty-two clone from ARDRA groups of pine humus forest soil were classified into 8 phyla:
-Proteobacteria, Acidobacteria, Bacteroides, Verrucomicrobia, Planctomycetes, and Gemmatomonadetes. According to PCA (Principal Component Analysis) based on 16S rRNA base sequence, there were three main groups of bacteria. All clone of Cluster I were originated from quercus humus forest soil, while 67% clone of Cluster II and 63% clone of Clusters III were separated from pine humus forest soil.
Distribution and Characteristics of Heterotrophic Plate Count Bacteria in Water Samples from Drinking Water Dispensers
Lee, Eun-Hwa ; Koh, Ji-Yun ; Kim, Jong-Seol ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 244~250
To evaluate bacteriological water quality, samples were taken from drinking water dispensers placed at S company (S-C) and U highschool (U-H) in Ulsan. The medians of heterotrophic plate counts (HPCs) were 53 CFU/ml for the 74 water samples of S-C and 80 CFU/ml for the 36 cold water samples of U-H, and 38% of the S-C and 42% of the U-H samples showed HPC bacterial concentrations higher than 100 CFU/ml. Coliform bacteria were detected from one sample of S-C. To determine the major source of bacterial contamination, water samples were taken daily for
days from the bottled water containers as well as the faucets of an experimental water dispenser. While the average HPCs in the bottled water containers were 33 CFU/ml for the first and 132 CFU/ml for the 2nd analysis, the HPC concentration in the cold water samples was 1,022 CFU/ml for the 2nd analysis. These results suggest that the majority of bacteria detected in the cold water samples were originated from the biofilms on the surface of water passages within the water dispensers. There was no significant increase in HPC bacterial concentrations within the bottled water container after installation on the water dispenser. We could isolate and tentatively identify 3 genera 6 species of Gram-positive and 7 genera 7 species of Gram-negative bacteria from the plate count agar plates of U-H samples. Among the isolates, 72% were observed as Gram-positive, and Micrococcus spp. was the most abundant with 54% of the total, followed by Sphingomonas paucimobilis with 16%. It appears that most of the HPC bacteria detected in water dispensers originate from indoor airborne bacteria, which may play important roles in the formation of biofilms on the surface of water passages within the water dispensers.
Isolation of Cyanobacteria Producing Microcystin from Lakes
Lee, Hee-Seon ; Oh, Kyoung-Hee ; Cho, Young-Cheol ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 251~257
Four potential microcystin-producing cyanobacteria were isolated from large reservoirs which act as sources of drinking water supply in Korea. Strain DC-2, YD-l, and YD-6 were closely related to Microcystis aeruginosa based on the analysis of l6S rRNA gene and mcyA gene sequences. mcyA gene sequence of YDS2-3, isolated from Yongdam Reservoir, was closed to that of M. aeruginosa, whereas l6S rRNA gene sequence was not related to the known sequences of microcystin-producing cyanobacteria indicating this strain can be a novel cyanobacterium belonging to the genus Microcystis. When mcyA gene sequences of isolated cyanobacteria were compared with the mcyA gene sequence library of two reservoirs, the sequence of DC-2 matched with the dominant ones.
Production of Antifungal Materials by Bacillus sp. Which Inhibit Growth of Phytophthora infestans and Fusarium oxysporum
Lee, Kang-Hyeong ; Song, Hong-Gyu ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 258~263
Late blight, one of the most important disease in many agricultural crops, is caused by Phytophthora infestans. Fusarium wilt is a vascular disease of many plants caused by Fusarium oxysporum. Some bacteria isolated from rhizosphere were screened for their ability to inhibit the growth of F. oxysporum and P. infestans. Productions of siderophore,
,3-glucanase, hydrogen cyanide and chitinase by 4 isolated strains were examined. Among them, Bacillus sp. RFO41 most effectively inhibited the growth of F. oxysporum. The highest productions of siderophore and
,3-glucanase were shown in the culture of Bacillus sp. RFO41. Bacillus strain PS2 was most effective against P. infestans. PS2 showed the highest production of chitinase and hydrogen cyanide. A significant relationship was shown between the antagonistic effects of isolates against F. oxysporum and P. infestans and their production level of siderophore,
,3-glucanase, hydrogen cyanide, and chitinase.
Characterization of Achlya bisexualis
-Amylase Expression in an Amylolytic Industrial Strain of Saccharomyces cerevisiae
Lee, Ok-Hee ; Lim, Mi-Hyeon ; Kim, Ji-Hye ; Ryu, Eun-Hye ; Ko, Hyun-Mi ; Chin, Jong-Eon ; Bai, Suk ;
The Korean Journal of Microbiology, volume 44, issue 3, 2008, Pages 264~269
To develop an amylolytic industrial yeast strain producing
-amylase, the BAMY gene encoding Achlya bisexualis
-amylase was constitutively expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) in an industrial strain of Saccharomyces cerevisiae. Yeast transformation was carried out by an integration system containing
-sequences as the recombination site. The integrative cassette devoid of bacterial DNA sequences was constructed that contains the BAMY gene and
-sequences. Industrial S. cerevisiae transformed with this integrative cassette secreted 45 kDa
-amylase into the culture medium. The
-amylase activity of the transformant was approximately 18.5-times higher than that of A. bisexualis. The multi-integrated BAMY genes in the transform ant were stable after 100 generations of growth in nonselective medium. Hydrolysis of soluble starch and various starches with the enzyme released maltose but not glucose or oligosaccharides.