• Radiation Oncology Journal
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• v.17 no.2
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• pp.151-157
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• 1999

Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.1977-1980
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• 2015

Objective: The Von Hippel-Lindau syndrome (VHLD), an inherited neoplastic syndrome predisposing to central nervous system hemangioblastoma (CNS), pheochromocytoma (PCC), renal cell carcinoma(RCC), retinal hemangioma (RA) and renal cysts, is caused by mutations or deletions of the VHL tumor-suppressor gene. To assess VHL genotype-phenotype correlations with function of pVHL a gene mutation analysis of members in a Chinese family with non-syndromic PCCs and individuals with apparently sporadic pheochromocytoma (ASP) was performed. Materials and Methods: DNA samples of 20 members from the Chinese family with non-syndromic PCCs and 41 patients with ASP were analyzed by polymerase chain reaction and direct sequencing, confirmed by Taqman probe. Results: Three novel mutations (H125P, 623(^TTTGTtG) and R120T) were identified in the Chinese family and in 3 among 41 ASP patients. The mutations were all located in exon 2 of VHL gene encoding ${\beta}$-domain of pVHL. The tumor type in H125P carriers and R120T carriers was VHL type 2C. And 623(^TTTGTtG) carriers presented VHL type 2B or type 2C. Conclusions: VHL gene abnormalities were identified in the Chinese family with non-syndromic PCCs and patients with APS, resulting in dysfunction of pVHL. H125P and R120T could be associated with VHL type 2C, while 623(^TTTGTtG) might be linked with VHL type 2B or type 2C. Not only is the genetic analysis helpful for early diagnosis and treatment of patients with VHLD, it is also benefitial for research intoVHLD pathogenesis.

• Journal of Dairy Science and Biotechnology
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• v.24 no.1
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• pp.19-28
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• 2006

This study was conducted to isolate antilisterial strains of the Bifidobacterium isolates from the infant feces. The bifidobacteria were isolated anaerobically on BL agar and screened for their inhibitory activity on the MRS-cysteine medium against three foodborne pathogens: Listeria monocytogenes, Bacillus cereus, and Staphylococcus aureus. Among the 52 bifidobacterial isolates, 5 strains(A24, Bl, B6, B10, and Bl2) were finally selected based on their stronger antilisterial activity against Listeria monocytogenes than other isolates tested. Morphologically, all the isolates were typically shown Y-and V-shaped under electron microscopic examination. Each isolate was primarily subjected to identification by a polymerase chain reaction(PCR) using a genus-specific primer designed for targeting the 16S rRNA gene sequence, and confirmed the primary identification data using an API-kit(Biomeriuex, France), commercially available product for identification based on biochemical and physiological traits. Of the isolates with antilisterial activity, strain A24 was finally confirmed as the Bifidobacterium longum A24.

• The Plant Pathology Journal
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• v.31 no.1
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• pp.25-32
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• 2015

Pseudomonas coronafaciens causes halo blight on oats and is a plant quarantine bacterium in many countries, including the Republic of Korea. Using of the certificated seed is important for control of the disease. Since effective detection method of P. coronafaciens is not available yet, PCR and TaqMan PCR assays for specific detection of P. coronafaciens were developed in this study. PCR primers were designed from the draft genome sequence of P. coronafaciens LMG 5060 which was obtained by the next-generation sequencing in this study. The PCR primer set Pc-12-F/Pc-12-R specifically amplified 498 bp from the 13 strains of P. coronafaciens isolated in the seven different countries (Canada, Japan, United Kingdom, Zimbabwe, Kenya, Germany, and New Zealand) and the nested primer set Pc-12-ne-F/Pc-12-ne-R specifically amplified 298 bp from those strains. The target-size PCR product was not amplified from the non-target bacteria with the PCR and nested primer sets. TaqMan PCR with Pc-12-ne-F/Pc-12-ne-R and a TaqMan probe, Pc-taqman, which were designed inside of the nested PCR amplicon, generated Ct values which in a dose-dependent manner to the amount of the target DNA and the Ct values of all the P. coronafaciens strains were above the threshold Ct value for positive detection. The TaqMan PCR generated positive Ct values from the seed extracts of the artificially inoculated oat seeds above 10 cfu/ml inoculation level. PCR and TaqMan PCR assays developed in this study will be useful tools to detect and identify the plant quarantine pathogen, P. coronafaciens.

• Journal of Microbiology
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• v.44 no.2
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• pp.155-161
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• 2006

Comparative analysis of microbial communities in a sequencing batch reactor which performed enhanced biological phosphorus removal (EBPR) was carried out using a cultivation-based technique and 16S rRNA gene clone libraries. A standard PCR protocol and a modified PCR protocol with low PCR cycle was applied to the two clone libraries of the 16S rRNA gene sequences obtained from EBPR sludge, respectively, and the resulting 424 clones were analyzed using restriction fragment length polymorphisms (RFLPs) on 16S rRNA gene inserts. Comparison of two clone libraries showed that the modified PCR protocol decreased the incidence of distinct fragment patterns from about 63 % (137 of 217) in the standard PCR method to about 34 % (70 of 207) under the modified protocol, suggesting that just a low level of PCR cycling (5 cycles after 15 cycles) can significantly reduce the formation of chimeric DNA in the final PCR products. Phylogenetic analysis of 81 groups with distinct RFLP patterns that were obtained using the modified PCR method revealed that the clones were affiliated with at least 11 phyla or classes of the domain Bacteria. However, the analyses of 327 colonies, which were grouped into just 41 distinct types by RFLP analysis, showed that they could be classified into five major bacterial lineages: ${\alpha},\;{\beta},\;{\gamma}-$ Proteobacteria, Actinobacteria, and the phylum Bacteroidetes, which indicated that the microbial community yielded from the cultivation-based method was still much simpler than that yielded from the PCR-based molecular method. In this study, the discrepancy observed between the communities obtained from PCR-based and cultivation-based methods seems to result from low culturabilities of bacteria or PCR bias even though modified culture and PCR methods were used. Therefore, continuous development of PCR protocol and cultivation techniques is needed to reduce this discrepancy.

• The Journal of the Korean Society for Microbiology
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• v.34 no.5
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• pp.491-500
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• 1999

A recent study showed that comparative sequence analysis of rpoB DNAs could reveal natural relationships in genus Mycobacterium [J Clin Microbial. 37 (6). 1999]. rpoB DNAs showed interspecies variation and intraspecies conservation. Based on these data, we developed polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) protocols which enable species differentiation in genus Mycobacterium. When this assay was applied to 24 clinical isolates identified as M. avium complex (MAC) by biochemical test, these were successfully differentiated into M. avium and M. intracellulare. These results were concordant with those obtained by 16s rDNA analysis. It is the first report that PCR-SSCP analysis of rpoB DNA could be used for species differentiation of MAC strains.

• Food Science and Biotechnology
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• v.17 no.1
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• pp.123-129
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• 2008

To isolate Bacillus cereus presenting at a level of 5 log CFU/g in kochujang, a primary dilution ($10^{-1}$) of kochujang was heated at $85^{\circ}C$ for 5 min. Two isolated strains Voges-Proskauer positive colony (KBC) and a negative colony (KBM) were identified as B. cereus and Bacillus mycoides, respectively, by biochemical test and 16S rDNA sequencing. $D_{100^{\circ}C}$-Values of KBC and KBM strains was 8.37 and 7.08 min, respectively. When spores of KBC strain were inoculated to kochujang at the level of 4-5 log spores/g, the number of spores was no significant difference (p<0.05) for each sample from 1 up to 60 day of aging. When kochujang was inoculated with 4 log spores/g and heated at $85^{\circ}C$ for 15 min, the number of spores was similar to that of unheated kochujang. Therefore, we estimated that B. cereus isolated from kochujang resistant on the heat treatment ($85^{\circ}C$, 15 min) and its heat resistance characteristics could be used to count the number in kochtjang.

• Advances in environmental research
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• v.2 no.3
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• pp.245-260
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• 2013

Biosorption represents a technological innovation as well as a cost effective excellent remediation technology for cleaning up radionuclides from aqueous environment. In the present study, a bacteria strain FB12 with high adsorption rate of uranium ion was isolated from the vicinity of the nuclear power plant. It was tentatively identified as Bacillus sp.FB12 according to the 16S rDNA sequencing. Efforts were made to further improve the adsorption rate and genetic stability by UV irradiation and UV-LiCl cooperative mutagenesis. The improved strain named Bacillus sp.UV32 obtains excellent genetic stability and a high adsorption rate of 95.9%. The adsorption of uranium U (VI) by Bacillus sp.UV32 from aqueous solution was examined as a function of metal ion concentration, cell concentration, adsorption time, pH, temperature, and the presence of some foreign ions. The adsorption process of U (VI) was found to follow the pseudo-second-order kinetic equation. The adsorption isotherm study indicated that it preferably followed the Langmuir adsorption isotherm. The thermodynamic parameters values calculated clearly indicated that the adsorption process was feasible, spontaneous and endothermic in nature. These properties show that Bacillus sp.UV32 has potential application in the removal of uranium (VI) from the radioactive wastewater.

• Journal of Environmental Science International
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• v.16 no.9
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• pp.1103-1109
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• 2007

The aim of this study was to isolate mesophilic chicken feather-degrading bacteria and to evaluate feather hydrolysate as alternative biofertilizer. Isolate RS7 was isolated from compost and identified as Bacillus pumilus according to API analysis and l6S rDNA sequencing analysis. Chicken feathers were completely degraded after 5 days of cultivation at $30^{\circ}C$. Feather hydrolysate treated by B. pumilius RS7 positively influenced Helianthus sannuus L. (sunflower) growth (e.g. growth rate, number and dry weight of leave, and flowering rate). These results suggest that feather hydrolysate prepared using B. pumilius RS7 could successfully be used as alternative biofertilizer, thereby reducing the environmental impact of feather waste from the poultry industry.

• Journal of Environmental Science International
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• v.17 no.4
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• pp.469-477
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• 2008

The aerobic photosynthetic bacterium, which produces bacterial carotenoids was isolated and identified from coastal marine environments. This bacterium was identified by 16S rDNA sequencing and designated as Erythrobacter longus SY-46. E. longus SY-46 was Gram negative and rod shape, and the optimal culture conditions were $25^{\circ}C$, pH 7.0, and 3.0% NaCl concentration, respectively. The carbon and nitrogen sources required for the optimal growth were lactose and tryptone, respectively. Fatty acid compositions of E. longus SY-46 were $C_{18:1}$(78.32%), v-linolenic acid($C_{18:3n9.12.15c}:3.83%$), margaric acid($C_{17:0}$: 3.38%), palmitic acid($C_{16:0}$: 3.07%), and docosahexaenoic acid($C_{22:6n3}$: 2.21%). In addition, E. longus SY-46 showed the characteristic absorption peaks of bacterial carotenoids(in the region of 450 to 480 nm) and bacteriochlorophyll(770 to 772 nm). Major carotenoids of E. longus SY-46 were polyhydroxylated xanthophylls such as fucoxanthin and zeaxanthin.