• Journal of Life Science
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• v.13 no.6
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• pp.856-864
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• 2003

The 16S rRNA gene is the most common gene in the phylogenetic analysis of procaryotes. However very high conservative of 16S rRNA has limitation in the discrimination of highly related organisms, hence other molecule was applied in this study and the result was compared with that of 16S rRNA. Three COGs (Clusters of Orthologous of protein) were only detected in 42 procaryotes ; transcription elongation facto. (COG0195), bacterial DNA primase (COG0358) and uridylate kinase (COG0528). Uridylate kinase gene was selected because of the similarity and one single copy number in each genome. Bacteria, belong to same genus, and Archaebacteria were same position with high bootstrap value in phylogenetic tree like the tree of 16S rRNA. However, alpha and epsilon Proteobcteria showed different position and Spirochaetales of Eubarteria was grouped together with Archaebacteria unlike the result of 16S rRNA. Uridylate kinase may compensate the problem of very high conservative of 16S rRNA gene and it would help to access more accurate discrimination and phylogenetic analysis of bacteria.

• Tuberculosis and Respiratory Diseases
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• v.45 no.6
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• pp.1178-1187
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• 1998

Background : Recently the incidence of tuberculosis is increasing in many countries and control of the disease is further threatened by the emergence of multi-drug resistant tuberculosis. So rapid detection of drug resistance is very important. Pyrazinamide (PZA) is a first-line chemotherapeutic agent for tuberculosis. Now in Korea, we perform PZase activity test instead of actual pyrazinamide susceptibility test for the detection of PZA resistant M. tuberculosis. Recently the pncA gene, encoding the PZase of M. tuberculosis, was completely sequenced. And it was reported that the mutation of pncA gene would be associated with PZA resistance of M. tuberculosis. Therefore we performed this study to evaluate the possibility for the rapid detection of PZA resistant M. tuberculosis using PCR-SSCP of pncA gene. Method : 44 cultured clinical isolates of M. tuberculosis, BCG Tokyo strain. BCG French strain, and one M. bovis isolate were studied. We used H37Rv as the reference strain, The PZase activity test was done at the reference laboratory of Korean Tuberculosis Institute. DNA was extracted by bead-beater method and 561 bp fragment including pncA gene was amplified by PCR. The PCR product were digested by BstB I enzyme. SSCP was done using MDE gel. Of the 44 strains of M. tuberculosis, 22 strains were PZase-positive and other 22 strains were PZase negative. Results : Of the 22 PZase positive strains, 18 strains(82%) showed the same mobility compared with that of H37Rv and 4(18%) showed different mobility. Of the 22 PZase-negative strains, 19(86%) strains showed the same mobility pattern compared with that of H37Rv and 3(14%) showed different mobility. Naturally PZA-resistant BeG-French strain, BCG-Tokyo strain, and one M. bovis isolate showed the same band pattern each other, but their mobility were different from that of H37Rv. The results of PZase activity test and PCR-SSCP of pncA of M. tuberculosis were statistically significantly correlated each other (p<0.01). Conclusion : The PCR-SSCP after BstB I restriction of pncA gene of M. tuberculosis may be a useful method for the rapid detection of PZA-resistant M. tuberculosis.

• The Korean Journal of Pesticide Science
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• v.8 no.4
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• pp.288-298
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• 2004

Benzimidazole pesticide carbendazim that is effective against a wide range of fungal plant pathogens is a protective, eradicant, and systemic fungicide. For genetic toxicity evaluation of carbendazim on DNA, genes and chromosome, were investigated with chromosome aberration, bacterial reverse mutation, micronucleus test in mouse born marrow and DNA damage assay by single cell microgel electrophoresis. Substitution and frameshift mutation were not induce at variable concentration of carbendazim on Ames test with or without rat liver microsomal activation. For the result of chromosome aberration test, numerical changes of chromosome were detected at the concentrations higher than $4.0{\mu}g/m{\ell}$, but structural aberration was not induced. Positive control, Mitomycin-C and captafol made a structural aberration, but numerical change of chromosome did not appear. In the micronucleus test for mouse born marrow, carbendazim was negative, but was weak positive in DNA damage assay by single cell microgel electrophoresis because of increased DNA moving length of 20% to control.

• Korean Journal of Environmental Agriculture
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• v.32 no.2
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• pp.148-154
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• 2013

BACKGROUND: Genetically modified(GM) trigonal cactus(Hylocereus trigonus Saff.) contained a coat protein gene of cactus virus X (CVX), which conferred resistance to the virus, phosphinothricin acetyltransferase (bar) gene, which conferred herbicide resistance, and a cauliflower mosaic virus 35S promoter (CaMV 35S). This study was conducted to evaluate the possible impact of GM trigonal cactus cultivation on the soil microbial community. METHODS AND RESULTS: Microorganisms were isolated from the rhizosphere of GM and non-GM trigonal cactus cultivation soils. The total numbers of bacteria, and actinomycete in the rhizosphere soils cultivated GM and non-GM trigonal cactus were similar to each other, and there was no significant difference. Dominant bacterial phyla in the rhizosphere soils cultivated with GM and non-GM trigonal cactus were Proteobacteria, Uncultured archaeon, and Uncultured bacterium. The denaturing gradient gel electrophoresis (DGGE) profiles show a similar patterns, significant difference was not observed in each other. DNA was isolated from soil cultivated GM and non-GM trigonal cactus, we analyzed the persistence of the inserted gene by PCR. Amplification of the inserted genes was not observed in the soil DNA, which was collected after harvest. CONCLUSION(S): This result suggests that the GM trigonal cactus cultivation does not change significantly the microbial community.

• Korean Journal of Environmental Agriculture
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• v.32 no.2
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• pp.95-101
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• 2013

BACKGROUND: Rice (Oryza sativa) is the most important staple food of over half the world's population. This study was conducted to evaluate the possible impact of transgenic rice cultivation on the soil microbial community. METHODS AND RESULTS: Microorganisms were isolated from the rhizosphere of GM and non-GM rice cultivation soils. Microbial community was identified based on the culture-dependent and molecular biology methods. The total numbers of bacteria, fungi, and actinomycete in the rhizosphere soils cultivated with GM and non-GM rice were similar to each other, and there was no significant difference between GM and non-GM rice. Dominant bacterial phyla in the rhizosphere soils cultivated with GM and non-GM rice were Actinobacteria, Firmicutes, and Proteobacteria. The microbial communities in GM and non-GM rice cultivated soils were characterized using the denaturing gradient gel electrophoresis (DGGE). The DGGE profiles showed similar patterns, but didn't show significant difference to each other. DNAs were isolated from soils cultivating GM and non-GM rice and analyzed for persistence of inserted gene in the soil by using PCR. The PCR analysis revealed that there were no amplified protox gene in soil DNA. CONCLUSION(S): These data suggest that transgenic rice does not have a significant impact on soil microbial communities, although continued research may be necessary.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.121-129
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• 1996

Multiplication of Herpes simplex virus type-1 was observed by electronmicroscopy, a gene library of the genome was constructed and thymidine kinase gene was cloned. Vero cells infected with the virus were lysed 48 h p.j. and multinucleated giant cells were observed approximately at 72 h p.i. The nucleocapsids were observed in nuclei and cytoplasm, and the assembled nucleocapsids were budded out through the vacuole and cytoplasmic membranes, and then virions were released from the cells. HSV-1 genome DNA was digested with BamHI and BglII enzymes and then the gene library of the genome fragments were constructed. The BamHI cleaved the genome DNA into twenty-seven fragments in the range of 1.1 - 14 kb, and BglII cleaved the genome DNA into sixteen fragments in the range of $4.5{\sim}20.1\;kb$. The pHLA-12 and pHLB-4 recombinant plasmids were contained TK gene by Southern blot analysis. The molecular sizes of the fragments which contained the TK gene were 3.74 in pHLA-12 and 6.41kb in pHLB-4 recombinant plasmid, respectively.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.323-328
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• 2017

Angiogenesis is a process including members of the angiogenic factors. In particular, fibroblast growth factor 2 (FGF2) is considered the most potent angiogenic factor because it promotes cell proliferation and tube formation. A recent study reported that fucoidan derived from marine plant potentiated FGF-2 induced tube formation in human endothelial cells. On the other hand, the molecular mechanisms involved in the angiogenic activity of fucoidan and FGF2 are unknown. In this study, a fucoidan treatment promoted angiogenesis induced by FGF2. The effects of fucoidan on FGF2-induced angiogenesis were confirmed by a proliferation assay using a CellTiter96 Aqueous One solution after a treatment with fucoidan and FGF2. The tube formation and wound healing assay for the angiogenic activity were also confirmed. Reverse transcription PCR showed a change in the mRNA of vascular endothelial growth factor-A (VEGF-A), intercellular adhesion molecule-1 (ICAM-1), matrix metallopeptidase9 (MMP9), and the signal transducer and activator of transcription3 (STAT3). In summary, the Fucoidan/FGF2 treatment induced an increase in cell proliferation, improved the tube formation and wound healing activity, and altered the STAT3, VEGF-A, ICAM-1, and MMP9 mRNA expression levels. Further research will be needed to provide a scientific explanation in terms of cell-signaling and confirm the present findings.

• Journal of Life Science
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• v.33 no.8
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• pp.605-611
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• 2023

Recently, sporadic cases of human infection by genetic reassortants of H7Nx influenza A viruses have been reported; such viruses have also been continuously isolated from avian species. In this study, A/wild bird/South Korea/sw-anu/2023, a novel reassortant of the H7N1 avian influenza virus, was analyzed using full-genome sequencing and molecular characterization. Phylogenetic analysis showed that A/wild bird/South Korea/sw-anu/2023 belonged to the Eurasian lineage of H7Nx viruses. The polymerase basic (PB)2, PB1, polymerase acidic (PA), and nucleoprotein (NP) genes of these viruses were found to be closely related to those of avian influenza viruses isolated from wild birds, while the hemagglutinin (HA), neuraminidase (NA), matrix (M), and nonstructural (NS) genes were similar to those of avian influenza viruses isolated from domestic ducks. In addition, A/wild bird/South Korea/sw-anu/2023 also had a high binding preference for avian-specific glycans in the solid-phase direct binding assay. These results suggest the presence of a new generation of H7N1 avian influenza viruses in wild birds and highlight the reassortment of avian influenza viruses found along the East Asian-Australasian flyway. Overall, H7Nx viruses circulate worldwide, and mutated H7N1 avian viruses may infect humans, which emphasizes the requirement for continued surveillance of the H7N1 avian influenza virus in wild birds and poultry.

• YAKHAK HOEJI
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• v.36 no.6
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• pp.582-590
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• 1992

S. pneumoniae recP mutant was compared with rec-8 mutant to identify whether they are the same gene or not by determining sensitivity to DNA damaging agents. recP and rec-8 mutant have almost same sensitivity to UV, ethylmethane sulfonate, and methylmethane sulfonate, suggesting that recP has the same function as the rec-8 gene in DNA repair.

• 대한핵의학회:학술대회논문집
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• 2002.11a
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• pp.35.1-35.1
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• 2002