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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
Journal Basic Information
Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 43, Issue 4 - Dec 2007
Volume 43, Issue 3 - Sep 2007
Volume 43, Issue 2 - Jun 2007
Volume 43, Issue 1 - Mar 2007
Selecting the target year
Potential Reproductive Toxicity Study of p53 Expressing Adenoviral Vector in Mice
Rhee, Gyu-Seek ; Kwack, Seung-Jun ; Kim, Soon-Sun ; Lee, Rhee-Da ; Seok, Ji-Hyun ; Chae, Soo-Young ; Chung, Soo-Youn ; Kim, Seung-Hee ; Lee, Seung-Hoon ; Park, Kui-Lea ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 151~158
The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with
PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.
Development of Hepatitis C Virus (HCV) Genome-Targeting Hammerhead Ribozyme Which Activity Can Be Allosterically Regulated by HCV NS5B RNA Replicase
Lee, Chang-Ho ; Lee, Seong-Wook ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 159~165
For the development of basic genetic materials for specific and effective therapeutic approach to suppress multiplication of hepatitis C virus (HCV), HCV internal ribosome entry site (IRES)-targeting hammerhead ribozyme which activity is allosterically regulated by HCV regulatory protein, NS5B RNA replicase, was developed. The ribozyme targeted most effectively to +382 nucleotide (nt) site of HCV IRES RNA. The allosteric ribozyme was designed to be composed of sequence of RNA aptamer to HCV NS5B, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding NS5B to the aptamer, and sequence of ribozyme targeting +382 nt of HCV IRES. Noticeably, we employed in vitro selection technology to identify the most appropriate communication module sequence which can induce ribozyme activity depending on the US5B protein. We demonstrated that the ribozyme was nonfunctional either in the absence of any proteins or in the presence of control bovine serum albumin. In sharp contrast, the allosteric ribozyme can induce activity of cleavage reaction with HCV IRES RNA in the presence of the HCV NS5B protein. This allosteric ribozyme can be used as lead compound for specific and effective anti-HCV agent, tool for highthroughput screening to isolate lead chemicals for HCV therapeutics, and ligand for biosensor system for HCV diagnosis.
Cloning of tlrD, 23S rRNA Monomethyltransferase Gene, Overexpression in Eschepichia coli and Its Activity
Jin, Hyung-Jong ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 166~172
ERM proteins transfer the methyl group to
in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confer the antibiotic resistance on micro-organisms ranging from antibiotic producers to pathogens and are classified into monomethyltransferase and dimethyltransferase. To investigate the differences between mono- and dimethyltransferase, tirD, a representative monomethylase gene was cloned in Escherichia coli from Streptomyces fradiae which contains ermSF, dimethylase gene as well to overexpress the TlrD for the first time. T7 promoter driven expression system successfully overexpress tlrD as a insoluble aggregate at
accumulating to around 55% of the total cell protein but unlike ErmSF, culturing at temperature as low as
did not make insoluble aggregate of protein into soluble protein. Coexpression of Thioredoxin and GroESL, chaperone was not helpful in turning into soluble protein either as in case of ErmSF. These results might suggest that differences between mono- and dimethylase could be investigated on the basis of the characteristics of protein structure. However, a very small amount of soluble protein which could not be detected by SDS-PAGE conferred antibiotic resistance on E. coli as in ErmSF which was expected from the activity exerted by monmethylase in a cell.
Epidemiological Characterization of Influenza Virus Isolated from Acute Respiratory Illness in Busan, 2004-2005
Cho, Kyung-Soon ; Park, Sun-Mee ; Kim, Seang-Joon ; Jung, Myung-Ju ; Lee, Joo-Yeon ; Kang, Chun ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 173~178
The occurrence of acute respiratory infections caused by the influenza virus are particularly high during the winter season in Busan, Korea. In 2004 and 2005, a study of the rate of occurrences of the influenza virus was conducted. The results reveal that in 2004, of the 1,869 people with an acute respiratory infection that 154 (8.2%) people were infected by the influenza virus. In 2005, of the 1,579 people infected with an acute respiratory infection that 19 people (1.2%) were infected with the influenza virus. The study shows a decrease in the numbers of an influenza virus infection from 2004 to 2005. Data was collected by inspecting throat swabs and nasal discharge from those with an acute respiratory infection. Further inspection of the throat swab and nasal discharge from the infected individuals during 2004 and 2005 study show the occurrence of the different types of influenza virus in the population: 6 cases (3.5%) of influenza type A/H1N1, 129 cases (74.5%) of A/H3N2, and 38 cases (22.0%) of type B. The study conducted in 2004 and 2005 reveal that children between the ages of two and five were more likely to be infected than any other age group. In the study, about 62.2% of the infected individuals were between two and five years old. The detection rates between males and females are similar. However, it is notable that females are slightly more likely to develop an acute respiratory infection caused by the influence virus compared to their male counterparts.
Detection of Ralstonia solanacearum with Nested PCR and DNA Enzyme-Linked Immunosorbent Assay
Ko, Young-Jin ; Cho, Hong-Bum ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 179~185
In this study, we used the method of guanidin isothiocyanate and boiling with Chelex-100 resin to extract genomic DNA of Ralstonia solanacearum from soil. It is more efficient than general protocols to remove inhibitory compounds in soil and R. solanacearum on. Then, we applied polymerase chain reaction and DNA enzyme-linked immunosorbent assay (ELISA) to identify and detect pathogen. The fliC gene of R. solanacearum was selected for specific detection of pathogen and primer sets were designed. Among the primer sets, two specific and sensitive primer sets, RsolfliC(forward: 5-GAACGCCAACGGTGCGAACT-3 and reverse; 5-GGCGGCCTTCAGGGAGGTC-3, designed by J.
et al.) and RS_247 (forward: 5-GGCGGTCTGTCGGCRG-3 and reverse; 5-CGGTCGCGTTGGCAAC-3 designed by this study), were designed to perform nested PCR. Nested PCR primer was labeled with biotin for hybridization between nested PCR product and probe to analyze with DNA ELISA.
Survey on the Status of Microbial Contamination of Chicken Meats Collected from Poultry Processing Plants in Nationwide
Woo, Yong-Ku ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 186~192
This study was conducted to survey the hygienic status of chicken meats on the microbial levels, which were collected from poultry processing plants located in the local provinces in nationwide including the JeJu island (n=15) in 1997. In particular, Salmonella spp., Campylobacter jejuni, and Listeria monocytogenes, which were retarded as one of the most important entero-pathogens relating to food home illness from poultry, were investigated on their isolation frequency including the other pathogens related on the food-borne illness. A total of 115 processed chickens were submitted on the present study. In general, the bacterial contamination frequency showed more or less lower
than those of sold on the retail and super markets and department stores because of lacking of cross-contamination incidences, depending on the total cells, Coliforms and Staphylococcal cells count. While, Salmonella species, Campylobacter jejuni, Listeria monocytogenes, and coagulase positive Staphylococcus aureus isolation frequency of chicken meats from slaughter houses were 58.3%, 37.4%, 43.5%, and 30.4%, in order. But the present microbial isolation data were a little lower levels than those of sold on the retail and super markets and famous department stores in Seoul and GyeongGi province at the same period. It seemed that the cross-contamination problems (including the human, environmental and instrumental factors) during the marketing stage (after the last processing procedure; rinsing step) had the major roles on the increasing of the microbial contamination frequency on the chicken meats after the slaughter houses.
Isolation and Characterization of Airborne Bacteria and Fungi in Indoor Environment of Elementary Schools
Kim, Na-Yeong ; Kim, Young-Ran ; Kim, Min-Kyu ; Cho, Du-Wan ; Kim, Jong-Seol ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 193~200
Indoor airborne bacterial and fungal concentrations were examined at classrooms and corridors of 3 elementary schools in Ulsan. Airborne microorganisms were collected with an impaction-type air sampler using plate count agar and dichloran rose bengal chloramphenicol agar. During the semester, concentrations of bacteria ranged
at classrooms and
at corridors, while those of fungi ranged
at classrooms and
at corridors. The bacterial concentrations showed larger variations between situations and schools compared to those of fungi. When airborne bacteria were isolated and identified, 84% were observed as Gram-positive, and Micrococcus spp. was the most abundant group with 61% of tested isolates, followed by genus Staphylococcus with 10%. The Micrococcus spp. isolates, of which 75% were identified as M. luteus, appeared to be from human origins. The protective pigments and substantial cell wall of Micrococcus may provide selective advantage for their survival in the air. We also isolated and identified 15 genera of filamentous fungi. The most common culturable fungi were Cladosporium, Aspergillus and Penicillium, and these 3 genera were 69% of fungal isolates. Genus Stachybotrys, of which S. chartarum is a well known producer of many potent mycotoxins, was also detected from one of the schools. further systematic studies are necessary with an emphasis on species identification and mycotoxin production of isolated fungal genera, including Aspergillus, Penicillium, and Stachybotrys.
Phylogenetic Characteristics of viable but Nonculturable Bacterial Populations in a Pine Mushroom (Tricholoma matsutake) Forest Soil
Kim, Yun-Ji ; Whang, Kyung-Sook ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 201~209
The CFDA (6-carboxyfluorescein diacetate) direct viable count method and plate count (PC) method using conventional nutrient broth (NB) medium and
diluted NB (DNB) medium were applied to samples collected from Mt. Yongdoo In Andong, in an effect to determine the number of living bacteria pine mushroom forest soil. The number of living bacteria determined via plate count in NB medium comprised
of the CFDA direct viable count, and the bacteria in the DNB medium comprised
. This result indicated that viable but nonculturable (VBNC) bacteria existed in the pine mushroom forest soil at a high percentage. The phylogenetic characteristics of the VBNC bacterial populations in the samples of pine mushroom (Tricholoma matsutake) forest soil were analyzed via the direct extraction of DNA and 16S rDNA-ARDRA. The 115 clones from pine mushroom forest soil were clustered into 31 different RFLP phylotypes by ARDRA. Based on the 16S rDNA sequences, the 31 ARDRA clusters were classified into 6 phylogenetic groups:
, Acidobacteria, Actinobacteria and Firmicutes. Among these bacterial populations, approximately 85% were classified as members of phylum Acidobacteria. The Acidobacteria phylum was shown to exist abundantly in the pine mushroom forest soil.
Comparison of the Phylogenetic Diversity of Humus Forest Soil Bacterial Populations via Different Direct DNA Extyaction Methods
Son, Hee-Seong ; Han, Song-Ih ; Whang, Kyung-Sook ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 210~216
The principal objective of this study was to analyze 16S rDNA-ARDRA of the humus forest soil via an improved manual method and an ISOIL kit on the basis of the UPGMA clustering of the 16S rDNA combined profile, 44 ARDRA clusters of 76 clones via the ISOIL kit method and 45 ARDRA clusters of 136 clones via the improved manual method. On the basis of the 16S rDNA sequences, 44 clones from the ARDRA clusters by the ISOIL kit were classified into 3 phyla :
, Acidobacteria and Actinobacteria. Using the improved manual method, the specimens were classified into 6 phyla : the
, Acidobacteria, Bacteroides, Verrucomicrobia, Planctomycetes and Gemmatomonadetes. As a result, the modified manual method indicated greater phylogenetic diversity than was detected by the ISOIL kit. Approximately 40 percent of the total clones were identified as
and 30 percent of the total clones were
and assigned to dominant phylogenetic groups using the ISOIL kit. Using the modified manual method, 41 percent of the total clones were identified as Acidobacteria and 28 percent of total clones were identified as
and assigned to dominant phylogenetic groups.
Isolation and Identification of Wild Yeast and Its Use for the Production of Grapewine
Kim, Jung-In ; Lee, Nam-Keun ; Hahm, Young-Tae ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 217~221
The domestic cultured Campbell's Early and Geubong grapes were fermented far the production of red wines with the isolated wild yeast Saccharomyces cerevisiae IJ850. For the isolation of wild yeast, Geubong and Campbell's Early grapejuices were naturally fermented at room temperature for 6 days without adding stater culture. The strain isolated from Geubong which has 1.8 times higher fermentative ability than the strains isolated Campbell Early was selected. The selected strain was identified by using 26S rDNA sequencing. The strain showed 99.7% of similarity with Saccharomyces cerevisiae and thus identified as Saccharomyces cerevisiae IJ850. It was investigated the fermentative ability as the start culture. For the production of grapewine, the final sugar concentrations of grapejuices were adjusted to the
with anhydrous glucose. The grapejuices were fermented at room temperature for 10 days in the air-locked bottles filled with
gas. The final yield and alcohol concentration of Campbell's Early and Geubong grapewines fermented with the isolated wild yeast were 80.8%, 11.0% and 87.8%, 13.0%, respectively. Between the isolated wild yeast S. cerevisiae IJ850 and the commercial yeast S. cerevisiae EC1118, total acidities of grapewines produced with wild yeast were lower than those produced with the commercial yeast. The pH values and the values of color analysis of grapewines produced with both strains were similar. The total phenol contents of campbell's Early and Geubong wines produced with the isolated yeast and the commercial yeast were obtained in the range of 75 to 125mg/L. In conclusion, S. cerevesiae IJ850 isolated from the domestic cultured Geubong grape is able to use to produce grapewines as stater culture.
Isolation and Charcterization of Micrococcus sp. HJ-19 Secreting Extracellular Protease
Cha, In-Tae ; Oh, Yong-Sik ; Roh, Dong-Hyun ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 222~226
Proteases are degradative enzymes which hydrolyze a peptide bond between amino acids and they are abundantly applied to commercial field. In order to screen new source of pretense, bacteria secreting extracellular pretense were isolated by enrichment culture from deep sea water samples of East Sea, Korea. A bacterium, named as HJ19, showed the best growth and the largest clear zone in plates supplemented skim milk at
. The partial DNA sequence analysis of the 16S rRNA gene, phenotypic tests and morphology identified that this strain was In genus Micrococcus. The strain HJ19 could not grow at
but it started growth and showed pretense activity at
. The optimal growth was at
and the maximal protease activity at
was about 480unit/ml.
Screening of the Natural Plant Extracts for the Antimicrobial Activity on Dental Pathogens
Baek, Dong-Heon ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 227~231
Many of antimicrobial components used in the mouthwash have disadvantages such as coloring of periodontal tissue and homing sensation. To find the more biocompatible antimicrobial agent, we have screened the 2,125 kinds of natural plant extracts obtained from by the Plant Extract Bank (Daejeon, Korea). The antimicrobial activity was determined by spectrophotometric growth inhibition assay for the major dental pathogens. For the Porphyromonas gingivalis, 19 plant extracts had an antimicrobial activity, 17 plant extracts for the Prevotella intermedia, 45 plant extracts for the Haemophilus actinomycetemcomitans and 85 plant extracts for the Streptococcus mutans. Among these effective extracts, 8 kind of natural plant extracts had an antimicrobial activity for more than 3 species of dental pathogens. In our experiment, Cudrania tricuspidata, Morus bombycis and Mallotus japonicas have antibacterial effects on the all kinds of major dental pathogens. Therefore these plant extracts have a possibility to be a candidate for major antibacterial components in dental products.
Identification of the Nitrifying Archaeal Phylotype Carrying Specific amoA Gene by Applying Digital PCR
Park, Byoung-Jun ; Park, Soo-Je ; Rhee, Sung-Keun ;
The Korean Journal of Microbiology, volume 43, issue 3, 2007, Pages 232~235
Mesophilic Crenarchaeota have been known to be predominant among ammonia-oxidizing microorganisms in terrestrial and marine environments. In this study, we determined the archaeal phylotypes carrying specific amoA by combining digital PCR and multiplex-nested PCR. Analysis of samples in which amoA and 16S rRNA gene were amplified showed that amoA gene diversity was relatively higher than that of 16S rRNA gene. Nitrifying archaeal group I.1a was dominant over I.1b group of crenarchaota and euryarchaeota. This approach could be applied for interrelating a functional gene to a specific phylotype in natural environments.