• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1503-1509
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• 2014

With the purpose of facilitating the process of stable strain generation, a shuttle vector for integration of genes via a double recombination event into two ectopic sites on the Sulfolobus acidocaldarius chromosome was constructed. The novel chromosomal integration and expression vector pINEX contains a pyrE gene from S. solfataricus P2 ($pyrE_{sso}$) as an auxotrophic selection marker, a multiple cloning site with histidine tag, the internal sequences of malE and malG for homologous recombination, and the entire region of pGEM-T vector, except for the multiple cloning region, for propagation in E. coli. For stable expression of the target gene, an ${\alpha}$-glucosidase-producing strain of S. acidocaldarius was generated employing this vector. The malA gene (saci_1160) encoding an ${\alpha}$-glucosidase from S. acidocaldarius fused with the glutamate dehydrogenase ($gdhA_{saci}$) promoter and leader sequence was ligated to pINEX to generate pINEX_malA. Using the "pop-in" and "pop-out" method, the malA gene was inserted into the genome of MR31 and correct insertion was verified by colony PCR and sequencing. This strain was grown in YT medium without uracil and purified by His-tag affinity chromatography. The ${\alpha}$-glucosidase activity was confirmed by the hydrolysis of $pNP{\alpha}G$. The pINEX vector should be applicable in delineating gene functions in this organism.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.287-296
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• 2007

A Lactobacillus paraplantarum strain producing a bacteriocin was isolated from kimchi using the spot-on-the lawn method and named L. paraplantarum C7 [15]. The bacteriocin, paraplantaricin C7, was found to inhibit certain Lactobacillus strains, including L. plantarum, L. pentosus, and L. delbrueckii subsp. lactis. It also inhibited Enterococcus faecalis, yet did not inhibit most of the other LAB (lactic acid bacteria) tested. The maximum level of paraplantaricin C7 activity was observed under the culture conditions of $25^{\circ}C$ and a constant pH of 4.5. Paraplantaricin C7 retained 90% of its activity after 10 min of treatment at $100^{\circ}C$ and remained stable within a pH range of 2-8. Based on a culture supernatant, paraplantaricin C7 was purified by DEAE-Sephacel column chromatography and $C_{18}$ reverse-phase HPLC. SDS-PAGE and activity staining were then conducted using the purified paraplantaricin C7, and its molecular mass determined to be about 3,800 Da. The 28 N-terminal amino acids from the purified paraplantaricin C7 were determined, and the structural gene encoding paraplantaricin C7, ppnC7, was cloned by PCR using degenerate primers based on the N-terminal amino acid sequence. The nucleotide sequences for ppnC7 and other neighboring orfs exhibited a limited homology to the previously reported plantaricin operon genes. Paraplantaricin C7 is a novel type II bacteriocin containing a double glycine leader sequence.

• Mycobiology
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• 제44권4호
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• pp.283-290
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• 2016

A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeast-malt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.

• 한국어병학회지
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• 제23권2호
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• pp.177-187
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• 2010

The Interferon stimulated gene 15 (ISG15) is strongly induced in many cell types by IFNs, viral infections, and double-stranded RNA (poly I:C). The ISG15 homologue cDNA was isolated from the rock bream LPS stimulated leukocyte cDNA library. The rock bream ISG15 homologue was found to consist of 833 bp encoding 157 amino acid residues. Compared with other known ISG15 peptide sequences, the most conserved regions of the rock bream ISG15 peptide were found to be the tandem ubiquitin-like domains and a C-terminal LRLRGG conjugating motif, characteristic of mammalian and non-mammalian ISG15 proteins. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the ISG15 sequence of rock bream and that of Atlantic salmon, Atlantic cod, northern snake head, black rockfish and olive flounder. The expression of the rock bream ISG15 molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following poly I:C stimulation, with a peak at 3 h post-stimulation. The rock bream ISG15 gene was predominantly expressed in the PBLs, spleen and gill.

• 한국정보통신학회논문지
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• 제13권4호
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• pp.805-812
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• 2009

기계 학습을 응용한 많은 침입 탐지 시스템들에서 n-그램 접근 방법이 사용되고 있다. 그러나, n-그램 접근방법은 확장이 어렵고, 주어진 시퀀스에서 획득한 n-그램들이 서로 겹치는 문제들을 가지고 있다. 본 연구에서는 이러한 문제들을 해결하기 위해, 일반화된 k-절단 서픽스트리 (generalized k-truncated suffix tree; k-TST) 기반의 n-그램 증강 나이브 베이스 (n-gram augmented naive Bayes) 알고리즘을 침입 시퀀스의 분류에 적용하여 보았다. 제 안된 시스템의 성능을 평가하기 위해 n-그램 특징들을 사용하는 일반 나이브 베이스 (naive Bayes) 알고리즘과 서포트 벡터 머신(support vector machines) 알고리즘과 본 연구에서 제안한 n-그램 증강 나이브 베이스 알고리즘을 호스트 기반 침입 탐지 벤치마크 데이터와 비교하였다. 공개된 호스트 기반 침입 탐지 벤치마크 데이터인 뉴 멕시코 대학(University of New Mexico)의 벤치마크 데이터에 적용해 본 결과에 따르면, n-그램 증강 방법이, n-그램이 나이브 베이스에 직접 적용되는 경우(예: n-그램 특징을 사용하는 일반 나이브 베이스), 생기는 독립성 가정에 대한 위배의 문제도 해결하면서, 동시에 더 정확한 침입 탐지기를 생성해냄을 알 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제25권1호
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• pp.98-108
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• 2015

Staphylococcus aureus is an important foodborne pathogen that causes diverse diseases ranging from minor infections to life-threatening conditions in humans and animals. To further understand its pathogenesis, the genome of the strain S. aureus FORC_001 was isolated from a contaminated food. Its genome consists of 2,886,017 bp double-stranded DNA with a GC content of 32.8%. It is predicted to contain 2,728 open reading frames, 57 tRNAs, and 6 rRNA operons, including 1 additional 5S rRNA gene. Comparative phylogenetic tree analysis of 40 complete S. aureus genome sequences using average nucleotide identity (ANI) revealed that strain FORC_001 belonged to Group I. The closest phylogenetic match was S. aureus MRSA252, according to a whole-genome ANI (99.87%), suggesting that they might share a common ancestor. Comparative genome analysis of FORC_001 and MRSA252 revealed two non-homologous regions: Regions I and II. The presence of various antibiotic resistance genes, including the SCCmec cluster in Region I of MRSA252, suggests that this strain might have acquired the SCCmec cluster to adapt to specific environments containing methicillin. Region II of both genomes contains prophage regions but their DNA sequence identity is very low, suggesting that the prophages might differ. This is the first report of the complete genome sequence of S. aureus isolated from a real foodborne outbreak in South Korea. This report would be helpful to extend our understanding about the genome, general characteristics, and virulence factors of S. aureus for further studies of pathogenesis, rapid detection, and epidemiological investigation in foodborne outbreak.

• 한국운동역학회지
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• 제17권4호
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• pp.115-125
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• 2007

Grould Reaction force(GRF) is important in human movements and GRF measurements are one of the most frequently used tool in biomechanical studies. In the studies of the golf swing motion, people refer to GRF as weight transfer. A successful golf swing motion requires many segments activation sequences which are controled by the nerve system. Due to the inter- and intra-individual variability of the human movement and the movement strategies, reliability of the measurements are important in human movement studies. Previous golf researches were based on group studies and certain events' values were analyzed. The purposes of this study were to determine the number of trials for the reliable golf swing GRF data collection, to reveal the variability level of the meaningful components of the golf swing GRF, and to classify the types of the golf swing GRF patterns. Twenty three male professional golfers($26.4{\pm}6.6$ years, $174.3{\pm}5.2\;cm$, $71.3{\pm}6.5\;kg$) signed an informed consent form prior to participation in this study. GRFs of driver swings were collected with Kistler 9285 force platform and 9865A amplifier, and calculated by the KwonGRF program(Visol, Korea). Sampling frequency was 1080 Hz. GRF data were trimmed from 1.5 s prior to the impact to 0.5 s after the impact. The number of trials for the reliable GRF collection was determined when the change in floating mean overs the 25 % of the standard deviation of that variable. Variabilities of the variables were determined by the coefficient of variation(CV) of 10 %. The types of GRF patterns were determined by visual inspection of the peak GRF shapes. The minimum number of trials for the reliable golf swing GRF data collection was five. Ten-trial seems more conservative. The value of the peak GRF was more reliable than the value of the impact GRF. The CV of the peak GRF and impact GRF were 7.4 %, 15.2 %, respectively. Because of the +/- sigh of the peak GRF appearance time, it was impossible to calculate CV of the peak GRF appearance time. Golf swing GRF patterns were classified as sing peak type, double peak type, and plateau peak type. This classification suggests the presence of the different golf swing weight transfer strategies.

• 대한바이러스학회지
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• 제28권3호
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• pp.275-285
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• 1998

Based on the geographic range and distribution of its rodent reservoir host, the European common vole (Microtus arvalis), Tula virus is likely to be widespread throughout Eurasia. Tula virus-infected voles have been captured in Central Russia, Austria, Czech and Slovak Republics, and the former Yugoslavia. Although serologic evidence for Hantaan (HTN) or Seoul (SEO) virus infection can be found in the vast majority of the more than 300 cases of hemorrhagic fever with renal syndrome (HFRS) occurring annually in Korea, approximately 4% of Korean patients with HFRS show a more than 4-fold higher antibody titer to Puumala (PUU) virus than to HTN or SEO virus by double-sandwich IgM ELISA, suggesting the existence of pathogenic Puumala-related hantaviruses in Korea. To further define the geographic distribution and genetic diversity of Tula virus in Eurasia and to investigate the existence of previously unrecognized Microtus-borne hantavirus in Korea, arvicolid rodents were captured in Lodz, Poland in 1995 and in Yunchon-kun, Kyungki-do during April to May, 1998. In addition, sera from 18 Korean HFRS patients who showed higher (or the same) antibody titer to Tula virus than HTN and SEO viruses were examined for hantavirus RNA by RT-PCR. Hantaviral sequences were not detected in any of the 18 patients or in 35 reed voles (Microtus fortis) in Korea. Alignment and comparison of a 208-nucleotide region of the S segment, amplified from lung tissues of two hantavirus-seropositive Marvalis captured in Poland, revealed $80.8{\sim}83.2%$ sequence similarity, respectively, with Tula virus strains from Central Russia and the Czech and Slovak Republics. Phylogenetic analysis indicated that the newfound Tula virus strains from Poland were closely related to other Tula hantaviruses from Eurasia.

• Journal of Animal Science and Technology
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• 제46권4호
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• pp.547-554
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• 2004

텔로미어란 진핵세포에 존재하는 DNA-protein 복합체로서 염색체의 말단부에 tandem repeated DNA 서열(TTAGGG)n과 특정 단백질로 구성되어 있으며 세포 분열이 진행함에 따라 이의 길이가 짧아지게 되고 일정 길이 이하가 되면 세포의 사망이 유기된다. 텔로미어의 역할은 게놈의 보호자로서 염색체의 안정성에 본질적으로 작용할고 감수분열시 상동염색체간의 접합에 주된 작용을 하는 것으로 알려져 있다. 본 연구는 소와 돼지의 성축에 대한 텔로미디어의 핵형과 각 염색체상 텔로미어의 양적 분포 양상을 제시하고자 Holstein과 Landrace를 공시하고 이들로부터 섬유아세포 배양으로 중기상을 획득한 다음 human telomeric DNA probe를 이용하여 형광접합보인법(FISH)으로 분석하였다. 실험 결과 소와 돼지의 모든 염색체의 양 말단부에 뚜렷한 텔로미어 프로브의 접합 양상을 발견할 수 있었다. 소의 경우 염색체들 간 텔로미디어의 양적 변이가 나타났으며 돼지의 경우는 모든 분석된 세포에서 특이적으로 6q1의 위치에 interstitial telomere가 존재하였다. 양적형광접합보인법(Q-FISH) 분석 결과 일부 염색체에서 한쪽 말단의 텔로미어 함량이 유의적으로 높은 것으로 분석되었고, 전체적으로 거의 모든 염색체에서 소, 돼지 공히 q-arm 말단주의 함유율이 p-arm 말단부에 비해 높은 것으로 나타났다. 또한, 염색체상 텔로미어의 상대적 함유율은 소가 돼지에 비해 높게 나타났으며, 전체 염색체 중 텔로미어의 상대적 함유율은 소, 돼지 모두 Y 염색체에서 가장 높았다.

• 콘크리트학회논문집
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• 제23권6호
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• pp.773-780
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• 2011

이 연구는 초대형 매스 구조물인 지하식 LNG 저장탱크의 바닥슬래브 및 측벽에 타설되는 매스 콘크리트의 재료특성, 배합조건, 양생조건 및 콘크리트의 타설시기와 초기온도, 외기온 등을 고려한 온도응력 해석 결과를 기술하였다. 해석 결과를 토대로 유해한 균열의 발생 가능성을 예측하고, 이를 방지하기 위한 방안을 제시하였다. 이 연구에서는 콘크리트의 단열온도 상승시험을 통하여 수화열 관리에 유리하다고 평가된 2종류(벨라이트 시멘트+석회석 미분말)의 최적배합조건을 선정하였다. 온도응력 해석의 결과에 따르면, 바닥슬래브 2단을 제외한 대부분의 분할타설 블록에서 관통균열지수가 1.2이상을 만족하였다. 바닥슬래브 2단의 경우 균열방지 대책으로 선행냉각 방안을 제시하였으며, 콘크리트의 초기온도를 $25^{\circ}C$ 범위에서 관리할 경우에는 대부분의 타설블록에서 관통균열지수 1.2이상을 만족하는 것으로 나타났다. 또한, 바닥슬래브의 경우, 표면균열지수가 1.2이상이기 때문에 양생조건을 준수하면 표면균열을 제어할 수 있으며, 측벽의 경우에도 표면균열지수가 1.0이상을 만족하기 때문에 균열의 수 및 폭을 제어할 수 있는 것으로 나타났다.