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HIV-1 Infection Causes Intracellular Expression of p53, Which Induces PKR Expression, Followed by Inhibition of HIV-1 Tat Activity.

Cheol Hee YOON ; Jin Young WOO ; Yong Soo BAE

Journal of Bacteriology and Virology.2004;34(2):157-166.

Few papers have reported that the HIV-1 replication was inhibited by p53 in the infected cells. However, the detail mechanism for the p53-medicated HIV-1 suppression has not yet been clearly demonstrated. In our previous report, we addressed that p53-mediated Tat suppression is very likely associated with PKR. In the present study, we found that the amounts of p53 in the HIV-1 infected cells increased over 10 times in the early stages of infection as much as those in normal cells. Particularly noteworthy is that the both exogenous p53 and endogenous p53 enhanced PKR expression in the transformed or treated cells, and the amounts of PKR induced by p53 were almost equivalent to those induced by interferon. In the PKR promoter studies using Ppkr-CAT (CAT reporter system under the control of PKR promoter), CAT activity induced by p53 was stronger than that by interferon, suggesting that the p53-mediated PKR expression might be more efficient than interferon under the control of PKR promoter. Co-immunoprecipitation experiments showed that PKR directly binds to Tat protein. We established eIF-2alpha dominant negative (S51A) Jurkat cells (JK/eIF2alpha-51A) to block the PKR-mediated cell cycle arrest or apoptosis. In the JK/eIF2alpha-51A cells, not only p53 but also PKR inhibited the Tat activity. Taken together, our results demonstrate that the HIV-1 infection induces p53, which enhances PKR expression by promoter activation, followed by the inhibition of the Tat activity, finally resulting in the inhibition of HIV-1 replication. Detail mechanisms for the PKR-mediated Tat inactivation are under investigation.
Animals ; Apoptosis ; Cats ; Cell Cycle Checkpoints ; Eukaryotic Initiation Factor-2 ; Gene Products, tat ; HIV-1* ; Humans ; Immunoprecipitation ; Interferons ; Jurkat Cells

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Application of Hantavirus N Protein Expressed by E. coli System.

Cheong Hee YU ; Jae Hwan NAM ; Teak Kyun CHOI ; Young Ran JU ; Chan Hee LEE ; Young Hack SHIN ; Keun Yong PARK

Journal of Bacteriology and Virology.2004;34(2):147-155.

Hantaan viruses cause haemorrhagic fever with renal syndrome (HFRS), resulting in severe morbidity and mortality in humans. The genome of Hantaan virus is composed of three segmented and single stranded negative sense RNA genome. In this study, we expressed nucleocapsid (N) proteins of Hantaan 76-118, Seoul 80-39 and Hantaan virus isolated in Korea (01-23) using E. coli system. These N proteins were fused with a thioredoxin protein for secretion of the expressed protein. The antigenicity of each expressed N proteins was examined in Western blot with sera from HFRS patients and normal controls. The expressed N proteins did not show any cross-reactivity with sera obtained from patients with leptospirosis and tsutsugamushi disease. These results suggest that our recombinant N proteins can be used for the development of diagnostic system to distinguish between HFRS and leptospirosis or tsutsugamushi.
Blotting, Western ; Fever ; Genome ; Hantaan virus ; Hantavirus* ; Hemorrhagic Fever with Renal Syndrome ; Humans ; Korea ; Leptospirosis ; Mortality ; Nucleocapsid ; RNA ; Scrub Typhus ; Seoul ; Thioredoxins

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Characterization and Detection of Enteric Viruses in Surface Water, Finished Water, Tap Water by Total Culturable Virus Assay (TCVA) Method.

Eun mi KO ; Seok Jea JANG ; Hee Jung KIM ; Eun suk HONG ; Yung Oh SHIN ; Sang Hyun KIM

Journal of Bacteriology and Virology.2004;34(2):137-145.

The information of species and quantity of enteric viruses in surface water, finished water, and tap water is important in helping understand the pathogenesis of viruses, providing information about health and hygiene, improving handling technique of drinking water, and establishing the standards of water quality. Using standard total culturable virus assay-most probable number (TCVA-MPN) method, we tried to detect infectious enteric viruses in surface water, finished water, and tap water samples that were collected and evaluated according to the information collection rule (ICR). The results obtained with TCVA method were compared to the results from both reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture-RT-PCR (ICC-RT-PCR) method. Five of 86 samples (5.8%) were positive as determined by the TCVA-MPN method. Two of 86 samples (2.3%) were positive for reovirus as determined by the RT-PCR and ICC-RT-PCR, and contained infectious reovirus. One of 86 samples (1.7%) was positive for coxsackievirus type B3 as determined by the RT-PCR and ICC-RT-PCR.
Drinking Water ; Hygiene ; Water Quality ; Water*

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Development of Multiplex PCR Detection of Blood-borne Viruses by Nucleic Acid Hybridization.

Hyun Ah YI ; Hong Ran KIM ; Hee Man LEE ; Min Seok PARK ; Seung Hwa HONG ; Si Hyung YOO ; Chan Hee LEE

Journal of Bacteriology and Virology.2004;34(2):129-136.

Polymerase chain reaction (PCR) has been used as a substitute for conventional serological methods in order to provide blood or blood products free from contaminating viruses and recently attempts have focused to detect 2 or 3 viruses by a single multiplex PCR (M-PCR) reaction. We were able to detect human immunodeficiency virus type 1 (HIV-1), hepatitis C virus (HCV), hepatitis B virus (HBV) and human cytomegalovirus (HCMV) simultaneously by a single M-PCR. However detection by gel electrophoresis of the products from M-PCR suffers from drawbacks such as low sensitivity and product sizes. Here we report enhanced detection systems of M-PCR based on nucleic acid hybridization with arrays built on membrane. Membrane array was manufactured by spotting appropriate probe DNAs on nylon membrane. Single or multiplex PCR was performed and the PCR products were labeled with DIG and allowed to hybridize with the membrane array. Results indicate that nonspecific hybridization was not observed for membrane DNA array. Additionally, membrane array method could detect small amount of viruses that were not detectable by conventional gel electrophoresis. At least 25-fold, and in some cases more than 125-fold increases in sensitivity was obtained with DNA array method. Thus, the nucleic acid hybridization with membrane array could be applied for the detection of M-PCR of viruses in blood or blood products.
Cytomegalovirus ; DNA ; Electrophoresis ; Female ; Hepacivirus ; Hepatitis B virus ; HIV-1 ; Humans ; Membranes ; Metrorrhagia ; Multiplex Polymerase Chain Reaction* ; Nucleic Acid Hybridization* ; Nylons ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction

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Distribution of Pathogenic Genes and Molecular Typing of Yersinia pseudotuberculosis Isolated from Spring Water in Seoul.

Mi Sun KIM ; Young Ki LEE ; Kyung Sig KIM ; Mi Ok SONG ; Mi Ja SHIM

Journal of Bacteriology and Virology.2004;34(2):121-128.

In order to investigate pathogenic genes and genetic relationships of Y. pseudotuberculosis strains, We isolated 9 strains of Y. pseudotuberculosis from 380 spring water sites in Seoul from 2000 to 2003. All isolates were distributed to the northeast area in Seoul. The isloates were analyzed for chromosomal virulence gene (inv) and plasmid-borne genes (yadA and lcrF) using PCR to assume pathogenicity. As a result, all isolates were positive for the inv gene, but only five isolates (55.6%) were positive for the yadA and lcrF genes. RAPD and PCR-ribotyping were tested and all isolates were grouped with 90% similarity. RAPD revealed 4 clusters and PCR-ribotyping revealed 2 clusters. The result of this experiments confirmed the view that RAPD had better powerful discrimination than PCR-ribotyping and RAPD typing was effective to distinguish between various strains of Y. pseudotuberculosis from spring water.
Discrimination (Psychology) ; Molecular Typing* ; Polymerase Chain Reaction ; Seoul* ; Virulence ; Yersinia pseudotuberculosis* ; Yersinia*

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Molecular Taxonomic Survey of Mycobacteria Clinical Isolates from Patients in Jeju Island by rpoB Gene Based Molecular Biological Methods.

Hong KIM ; Sun Hyun KIM ; Gill Han BAI ; Young Gil PARK ; Chang Yong CHA ; Yoon Hoh KOOK ; Bum Joon KIM

Journal of Bacteriology and Virology.2004;34(2):113-119.

The rpoB gene based sequencing analysis enabled not only the detection of rifampin resistant Mycobacterium tuberculosis, but also the differentiation of species in the genus Mycobacterium. In the present study, we applied the method to 68 isolates of M. tuberculosis (29 from initial treatment cases and 39 from recurrent cases) and 11 clinical isolates of nontuberculous mycobacteia (NTM) isolated from patients in Jeju island. Among rifampin resistant M. tuberculosis, two of 29 strains isolated from patients of initial cases (6.9%) and five of 39 strains isolated from patients of recurrent cases (12.8%) were confirmed to have rifampin resistant genotypes harboring mutations in rif r region of the rpoB gene. In NTM strains, M. fortuitum complex was the most frequently isolated species at the frequency of 54.5% (6/11).
Genotype ; Humans ; Mycobacterium ; Mycobacterium tuberculosis ; Rifampin ; Tuberculosis

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Analysis of the Cell Lysate Proteome of a Korean Mycobacterium tuberculosis Isolate K01 with H37Rv and H37Ra Strains.

Sung Weon RYU ; Sang Chan PARK ; Mun Nam BANG ; Sung Sik HAN ; Young Kil PARK ; Sue Nie PARK ; Young Soo SHIM ; Seongman KANG ; Gill Han BAI

Journal of Bacteriology and Virology.2004;34(2):107-112.

Despite recent economic prosperity, Korea still has high prevalence of tuberculosis. Molecular biologic characterization of Korean Mycobacterium tuberculosis strains might provide a deeper understanding of the forces contributing to the spread of tuberculosis in Korea. Therefore, we analyzed the cell lysate proteome of a representative Korean Mycobacterium tuberculosis isolate (K01) in comparison with laboratory reference strains H37Rv and H37Ra. Seven spots were strongly expressed only in K01 strain compared with M. tuberculosis H37Rv and H37Ra. Through continuous MALDI-MS analysis, these spots were identified as hypothetical protein Rv3849, secreted immunogenic protein Mpt64, Acetyl/propionyl-CoA Carbpxylase (AccD1), alkyl hydroperoxide reductase C (AhpC), N-acetylmuramyl-L-alanine amidase, a putative UDP glucose epimerase, and a transposase. A deeper study of these proteins may provide a clue in the development of effective new anti-tuberculosis vaccines against Korean M. tuberculosis isolates.
Korea ; Mycobacterium tuberculosis* ; Mycobacterium* ; Peroxiredoxins ; Prevalence ; Proteome* ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Transposases ; Tuberculosis ; UDPglucose 4-Epimerase ; Vaccines

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Glucan Production and Glucosyltransferase Gene Expression in the Samples of Caries-active and Caries-free Children.

Jin CHUNG ; Dong Hoon LEE ; Tae Sung CHUNG ; Shin KIM ; Jong Suk OH

Journal of Bacteriology and Virology.2004;34(2):101-106.

It is well known that the number of Streptococcus mutans is closely related with dental caries activity. In this study, the capability of glucan production and the glucosyltransferase (gtf) gene expression were examined in the saliva and plaque samples from caries-active and caries-free children. Bacterial cells from each sample were estimated by viable cell count on selective media. The capability of glucan production was determined by measuring the amount of glucan synthesized on orthodontic wires in a medium inoculated with S. mutans isolates from both groups. The mRNA expression for the gtfB and gtfD genes was examined by Fluorescent In Situ Hybridization (FISH). In accordance with previous reports, the numbers of mutans streptococci (MS) and Lactobacilli were higher in caries-active saliva and plaque than in caries-free samples. S. mutans isolated from caries-active samples produced a large amount of glucan, whereas S. mutans from caries-free samples produced little glucan on the orthodontic wires. Finally, the expression of the gtfB and gtfD genes in caries-active plaques was higher than in caries-free plaques. The results suggest that the glucan production and the gtf gene expression as well as the level of MS and lactobacilli should be higher in the samples of caries-active children than those of caries-free children.
Cell Count ; Child* ; Dental Caries ; Gene Expression* ; Humans ; In Situ Hybridization, Fluorescence ; Orthodontic Wires ; RNA, Messenger ; Saliva ; Streptococcus mutans

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Rapid Detection of Methicillin-Resistant Staphylococcus aureus by Multiplex PCR.

Ji Young MOON ; Eun Jung LEE ; Yung Bu KIM

Journal of Bacteriology and Virology.2004;34(2):91-100.

Staphylococcus aureus continues to be the main cause of surgical site infections. Recently, methicillin-resistant S. aureus (MRSA) has been known to be resistant to many kinds of antibiotics and causes the problem of neonatal nosocomial infection. Antibiotic sensitivity tests which have been routinely used to detect MRSA in the laboratory depend on the culture conditions. Therefore it is necessary to develop a new method based on a molecular biological technique in order to overcome these problems. We report the development of a multiplex PCR protocol for the diagnosis of staphylococcal infection. The protocol was designed to i) detect any staphylococcal species to the exclusion of other bacterial pathogens (based on primers corresponding to staphylococcal-specific regions of the 16S rRNA genes), ii) provide an indication of the likelihood that the Staphylococci present in the specimen are resistant to oxacillin (based on the amplication of the mecA gene, encoding penicillin-binding protein 2'(PBP-2'), which is known to confer resistance to the bacteriostatic action of methicillin). In this study, 67 S. aureus strains were isolated from the neonatal intensive care unit and general neonatal nursery at Pusan National University Hospital, Busan, Korea, between January and July 2003. Methicillin resistance was tested by the oxacillin disk diffusion method and the MIC method. We performed the multiplex PCR to amplify the mecA gene, encoding PBP-2'. We tested it by multiplex PCR and compared the results with the antimicrobial susceptibilities. Different results were obtained from 2 MRSA (4.65%), suggesting that the PCR method should be performed at the same time for a more accurate clinical test of MRSA
Anti-Bacterial Agents ; Busan ; Cross Infection ; Diagnosis ; Diffusion ; Infant, Newborn ; Intensive Care, Neonatal ; Korea ; Methicillin Resistance* ; Methicillin-Resistant Staphylococcus aureus* ; Multiplex Polymerase Chain Reaction* ; Nurseries ; Oxacillin ; Penicillin-Binding Proteins ; Polymerase Chain Reaction ; Staphylococcal Infections ; Staphylococcus aureus ; Thiram

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Seroepidemiological Survey of Aujeszky's Disease Virus in Wild Boar (Sus scrofa) and Raccoon Dogs (Nyctereutes procyonoides koreensis) in Korea.

Dong Kun YANG ; Jin Ju NAH ; Ha Hyun KIM ; Sung Suk CHOI ; Dong Jun AN ; Jong Taek KIM ; Kyoung Ae PARK ; Jae Young SONG

Journal of Bacteriology and Virology.2014;44(4):336-341. doi:10.4167/jbv.2014.44.4.336

Aujeszky's disease caused by Aujeszky's disease virus (ADV) is one of the most important diseases in the pig industry. In this study, we conducted a seroepidemiological survey of ADV in wild boars and raccoon dogs in South Korea. In total, 217 wild boar sera collected between March and August 2013, and 96 raccoon dogs between 2011 and 2012 were screened for the presence of antibodies against ADV. The sero-positive rates in wild boars and raccoon dogs tested for ADV were found to be 3.55% (8/225) and 0% (0/96), respectively. The presence of virus neutralization antibody titer against ADV means that small number of wild boars was infected with ADV and AD may be circulated continuously in Korean wild boar populations, and that wild boars may act as a potential reservoir of ADV. Therefore, to achieve the declaration of AD free, effective preventive measures to block transmission of AD should be taken to the wild boars.
Antibodies ; Herpesvirus 1, Suid* ; Korea ; Pseudorabies ; Raccoon Dogs* ; Sus scrofa*

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