• Journal of Microbiology and Biotechnology
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• 제21권4호
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• pp.421-429
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• 2011

For precise identification of a Lactobacillus K1 isolate, LC-MS/MS analysis of the putative surface layer protein was performed. The results obtained from LTQ-FT-ICR mass spectrometry confirmed that the analyzed protein spot is the surface layer protein originating from Lb. helveticus species. Moreover, the identified protein has the highest similarity with the surface layer protein from Lb. helveticus R0052. To evaluate the proteomic study, multilocus sequence analysis of selected housekeeping gene sequences was performed. Combination of 16S rRNA sequencing with partial sequences for the genes encoding the RNA polymerase alpha subunit (rpoA), phenylalanyl-tRNA synthase alpha subunit (pheS), translational elongation factor Tu (tuf), and Hsp60 chaperonins (groEL) also allowed to classify the analyzed isolate as Lb. helveticus. Further classification at the strain level was achieved by sequencing of the slp gene. This gene showed 99.8% identity with the corresponding slp gene of Lb. helveticus R0052, which is in good agreement with data obtained by nano-HPLC coupled to an LTQ-FT-ICR mass spectrometer. Finally, LC-MS/MS analysis of surface layer proteins extracted from three other Lactobacillus strains proved that the proposed method is the appropriate molecular tool for the identification of S-layer-possessing lactobacilli at the species and even strain levels.

• Journal of Microbiology and Biotechnology
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• 제30권2호
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• pp.259-270
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• 2020

Listeria monocytogenes is a gram-positive, facultative anaerobe food pathogen responsible for the listeriosis that mostly occurs during the low-temperature storage of a cold cut or dairy products. To understand the systemic response to a wide range of growth temperatures, L. monocytogenes were cultivated at a different temperature from 10℃ to 42℃, then whole cell proteomic analysis has been performed both exponential and stationary cells. The specific growth rate increased proportionally with the increase in growth temperature. The maximum growth rate was observed at 37℃ and was maintained at 42℃. Global protein expression profiles mainly depended on the growth temperatures showing similar clusters between exponential and stationary phases. Expressed proteins were categorized by their belonging metabolic systems and then, evaluated the change of expression level in regard to the growth temperature and stages. DnaK, GroEL, GroES, GrpE, and CspB, which were the heat&cold shock response proteins, increased their expression with increasing the growth temperatures. In particular, GroES and CspB were expressed more than 100-fold than at low temperatures during the exponential phase. Meanwhile, CspL, another cold shock protein, overexpressed at a low temperature then exponentially decreased its expression to 65-folds. Chemotaxis protein CheV and flagella proteins were highly expressed at low temperatures and stationary phases. Housekeeping proteins maintained their expression levels constant regardless of growth temperature or growth phases. Most of the growth related proteins, which include central carbon catabolic enzymes, were highly expressed at 30℃ then decreased sharply at high growth temperatures.

• Journal of Microbiology and Biotechnology
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• 제25권9호
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• pp.1410-1416
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• 2015

Wolbachia is an obligate symbiotic bacteria that is ubiquitous in arthropods, with 25-70% of insect species estimated to be infected. Wolbachia species can interact with their insect hosts in a mutualistic or parasitic manner. Sequence types (ST) of Wolbachia are determined by multilocus sequence typing (MLST) of housekeeping genes. However, there are some limitations to MLST with respect to the generation of clone libraries and the Sanger sequencing method when a host is infected with multiple STs of Wolbachia. To assess the feasibility of massive parallel sequencing, also known as next-generation sequencing, we used pyrosequencing for sequence typing of Wolbachia in butterflies. We collected three species of butterflies (Eurema hecabe, Eurema laeta, and Tongeia fischeri) common to Korea and screened them for Wolbachia STs. We found that T. fischeri was infected with a single ST of Wolbachia, ST41. In contrast, E. hecabe and E. laeta were each infected with two STs of Wolbachia, ST41 and ST40. Our results clearly demonstrate that pyrosequencing-based MLST has a higher sensitivity than cloning and Sanger sequencing methods for the detection of minor alleles. Considering the high prevalence of infection with multiple Wolbachia STs, next-generation sequencing with improved analysis would assist with scaling up approaches to Wolbachia MLST.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1635-1643
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• 2009

The aim of this study was to provide new insight into the mechanism whereby the housekeeping enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) locates to cell walls of Lactobacillus plantarum 299v. After purification, cytosolic and cell wall GAPDH (cw-GAPDH) forms were characterized and shown to be identical homotetrameric active enzymes. GAPDH concentration on cell walls was growth-time dependent. Free GAPDH was not observed on the culture supernatant at any time during growth, and provoked cell lysis was not concomitant with any reassociation of GAPDH onto the cell surface. Hence, with the possibility of cw-GAPDH resulting from autolysis being unlikely, entrapment of intracellular GAPDH on the cell wall after a passive efflux through altered plasma membrane was investigated. Flow cytometry was used to assess L. plantarum 299v membrane permeabilization after labeling with propidium iodide (PI). By combining PI uptake and cw-GAPDH activity measurements, we demonstrate here that the increase in cw-GAPDH concentration from the early exponential phase to the late stationary phase is closely related to an increase in plasma membrane permeability during growth. Moreover, we observed that increases in both plasma membrane permeability and cw-GAPDH activity were delayed when glucose was added during L. plantarum 299v growth. Using a double labeling of L. plantarum 299v cells with anti-GAPDH antibodies and propidium iodide, we established unambiguously that cells with impaired membrane manifest five times more cw-GAPDH than unaltered cells. Our results show that plasma membrane permeability appears to be closely related to the efflux of GAPDH on the bacterial cell surface, offering new insight into the understanding of the cell wall location of this enzyme.

• Fisheries and Aquatic Sciences
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• 제19권4호
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• pp.21.1-21.11
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• 2016

Background: The evaluation of suitable reference genes as normalization controls is a prerequisite requirement for launching quantitative reverse transcription-PCR (RT-qPCR)-based expression study. In order to select the stable reference genes in abalone Haliotis discus hannai tissues (gill and hepatopancreas) under heavy metal exposure conditions (Cu, Zn, and Cd), 12 potential candidate housekeeping genes were subjected to expression stability based on the comprehensive ranking while integrating four different statistical algorithms (geNorm, NormFinder, BestKeeper, and ${\Delta}CT$ method). Results: Expression stability in the gill subset was determined as RPL7 > RPL8 > ACTB > RPL3 > PPIB > RPL7A > EF1A > RPL4 > GAPDH > RPL5 > UBE2 > B-TU. On the other hand, the ranking in the subset for hepatopancreas was RPL7 > RPL3 > RPL8 > ACTB > RPL4 > EF1A > RPL5 > RPL7A > B-TU > UBE2 > PPIB > GAPDH. The pairwise variation assessed by the geNorm program indicates that two reference genes could be sufficient for accurate normalization in both gill and hepatopancreas subsets. Overall, both gill and hepatopancreas subsets recommended ribosomal protein genes (particularly RPL7) as stable references, whereas traditional housekeepers such as ${\beta}-tubulin$ (B-TU) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were ranked as unstable genes. The validation of reference gene selection was confirmed with the quantitative assay of MT transcripts. Conclusions: The present analysis showed the importance of validating reference genes with multiple algorithmic approaches to select genes that are truly stable. Our results indicate that expression stability of a given reference gene could not always have consensus across tissue types. The data from this study could be a good guide for the future design of RT-qPCR studies with respect to metal regulation/detoxification and other related physiologies in this abalone species.

• 한국생활과학회지
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• 제6권2호
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• pp.83-93
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• 1997

In order to adapt to the economic change with the rapid world change, the wise consumers should have positive cognizance of their roles and are requested to fulfil their duties as a consummer. Such capability of the consumer could be acquired from home, mass media and school, and specially the role of mother at home would have a great influence on their early children's consummer activities. The purpose of this study was to investigate the interrelationship of the consumer behavior between mother and their children. As a result of this study, the mother with higher educational background could manage effectively the allowance of the children. The mother with older age had a great influence on the consumer's role of the children and high record of the execution. Employed mother had higher level of execution for the consumer's rights than their count part. Accordingly, the consumer behavior of the mother will have an influence on the children's consumer behavior. Such a result showed the meaningful statistic relations(r=3.6, p<.001) for the economy activity of the children the home management and economy life of the mother, and the purchasing activity of the children including the execution of the consumer's right. In their regard, there was the significant statistic relations(r=19, p<.05) between the children's pocket money and the housekeeping management of the mother. In the meantime, it showed that another changing factor concerned with the consumer behavior of the mother have little influence the children's allowance.

• 정보관리학회지
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• 제31권3호
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• pp.331-351
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• 2014

이 연구는 OECD PISA에 참여한 우리나라 학생, 학부모와 학교장의 설문자료와 읽기 수학 과학소양의 성취도를 분석하여 학교도서관변인이 학업성취도에 미치는 영향력을 파악하고자 하였다. 이를 통해 학교도서관의 교육적 영향력과 책무성에 대한 기초자료를 얻고 학교도서관 교육을 개선하기 위한 시사점을 도출하고자 하였다. 분석결과, 첫째, 학생들이 독서를 즐겁게 생각할수록 학생들의 학업성취도가 높았다. 둘째, 가정에 책이 많고 이용할 수 있는 독서자원이 많을수록 학생들의 학업성취도가 높았다. 셋째, 인터넷을 사용하기 위해 학교도서관을 방문할수록 학업성취도가 낮았다. 넷째, 학교도서관 직원이 부족할수록 학생들의 학업성취도가 낮았다. 즉 학교도서관 직원이 학생들의 학업성취도에 긍정적인 영향을 미치고 있었다. 다섯째, 독서 사교육에 대한 경험은 읽기 과학소양성취도에 부정적인 영향을 미치고 있었다. 여섯째, 독서가 좋아서 학교도서관에서 대출하는 것과 국어교과 관련 학교도서관 수업은 학업성취도에 영향을 미치지 않는 것으로 나타났다.

• 대한약침학회지
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• 제19권1호
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• pp.7-15
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• 2016

Objectives: Malaria has been a major global health problem in recent times with increasing mortality. Current treatment methods include parasiticidal drugs and vaccinations. However, resistance among malarial parasites to the existing drugs has emerged as a significant area of concern in anti-malarial drug design. Researchers are now desperately looking for new targets to develop anti-malarials drug which is more target specific. Malarial parasites harbor a plastid-like organelle known as the 'apicoplast', which is thought to provide an exciting new outlook for the development of drugs to be used against the parasite. This review elaborates on the current state of development of novel compounds targeted againstemerging malaria parasites. Methods: The apicoplast, originates by an endosymbiotic process, contains a range of metabolic pathways and housekeeping processes that differ from the host body and thereby presents ideal strategies for anti-malarial drug therapy. Drugs are designed by targeting the unique mechanism of the apicoplasts genetic machinery. Several anabolic and catabolic processes, like fatty acid, isopenetyl diphosphate and heme synthess in this organelle, have also been targeted by drugs. Results: Apicoplasts offer exciting opportunities for the development of malarial treatment specific drugs have been found to act by disrupting this organelle's function, which wouldimpede the survival of the parasite. Conclusion: Recent advanced drugs, their modes of action, and their advantages in the treatment of malaria by using apicoplasts as a target are discussed in this review which thought to be very useful in desigining anti-malarial drugs. Targetting the genetic machinery of apicoplast shows a great advantange regarding anti-malarial drug design. Critical knowledge of these new drugs would give a healthier understanding for deciphering the mechanism of action of anti-malarial drugs when targeting apicoplasts to overcome drug resistance.

• 한국어류학회지
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• 제18권4호
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• pp.293-299
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• 2006

두툽상어 (Scyliorhinus torazame)부터 분리된 항산화 효소 Cu,Zn-superoxide dismutase (Cu,Zn-SOD 또는 SOD1)의 핵산 염기서열 및 추정 아미노산 서열을 대상으로 분자 계통학적 분석을 실시하였다. 종래 알려져 있는 척추동물의 Cu,Zn-SOD 서열들을 포함하여 neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) 및 Bayesian 분석 등을 포함한 다양한 계통 분석을 수행하였으며, 이를 통해 연골어류인 본 어종의 척추동물 분류군 내에서의 계통적 위치를 추정하고자 하였다. 다양한 분자 계통수로부터 얻어진 대부분의 consensus tree들에서 분석에 사용한 분류군들은 종래 알려진 분류학적 위치와 비교적 잘 일치하였고, 이중 두툽상어는 같은 연골어류종인 blue shark와 높은 유연관계를 나타내면서 보다 진화한 경골어류들과는 확연히 구분되는 분지를 형성하였다. 특히 핵산 염기서열을 바탕으로 한 neighbor-joining 분석에서 두툽상어는 경골어류와 양막동물에 비해 보다 원시형태의 척추동물 Cu,Zn-SOD 유전자의 한 형태를 보유하고 있는 것으로 나타났다.

• Genomics & Informatics
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• 제1권1호
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• pp.40-49
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• 2003

The genes related to specific events or pathways in bacteria are frequently localized proximate to the genome of their neighbors, as with the structures known as operon, but eukaryotic genes seem to be independent of their neighbors, and are dispersed randomly throughout genomes. Although cases are rare, the findings from structures similar to prokaryotic operons in the nematode genome, and the clustering of housekeeping genes on human genome, lead us to assess the genomic association of genes as functional subunits. We evaluated the genomic association of neighboring genes on chromosomes 4 and 5 of Arabidopsis thaliana with and without respectively consideration of the scaffold/matrix­attached regions (S/MAR) loci. The observed number of functionally identical bigrams and trig rams were significantly higher than expected, and these results were verified statistically by calculating p-values for weighted random distributions. The observed frequency of functionally identical big rams and trig rams were much higher in chromosome 4 than in chromosome 5, but the frequencies with, and without, consideration of the S/MAR in each chromosome were similar. In this study, a genomic association among functionally related neighboring genes in Arabidopsis thaliana was suggested.