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Isolation and Characterization of Bacillus Species Having Antifungal Activity Against Pathogens of Ginseng Damping Off (인삼모잘록병원균에 항균활성을 갖는 Bacillus 균의 분리 및 특성조사)

  • Park, Kyeong Hun;Park, Hong Woo;Lee, Seong Woo;Lee, Seung Ho;Myung, Kyung Sun;Lee, Sang Yeob;Song, Jaekyeong;Kim, Young Tak;Park, Kyoung Soo;Kim, Young Ock
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.380-387
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    • 2016
  • This study was performed to select potentially available biological control agent from soil bacteria for prevention of ginseng damping off. More than five hundred strains were isolated from ginseng rhizosphere soil. By testing antifungal activity, we have selected three soil bacteria strains and their ability to produce antibiotics and lytic enzymes such as cellulase, protease and pectate lyase was examined. Also, the presence of genes for biosynthesis of lipopeptide such as fengycin, bacillomycin D, surfactin, iturin A, and zwittermicin A was investigated in selected strains. All three strains produced cellulase, protease, and xylanase. Moreover, these strains had gene for biosynthesis of bacillomycin D, surfactin, and iturin A. ES1 and ES3 strains were identified Bacillus methylotrophucus and ES2 was confirmed Bacillus amyloliquefaciens using phylogenetic analysis on the basis of 16S rRNA gene sequences. In field test, control value of ES1, ES2 and ES3 treatment was 32.4%, 46.8% and 36.7%, respectively. This results indicate that antagonistic microbes with high ability of antifungal and lytic enzyme activity can be used as a useful biological control agent to control ginseng damping off.

Comparison of the Bacterial and Fungal Colonies from Rana dybowskii which Collected from Inside and Outside Frog Farms and Identification of the Bacteria from the Tadpoles (개구리 증양식장 내·외부에서 채집된 북방산개구리(Rana dybowskii)로부터 검출된 세균과 곰팡이 콜로니 수의 비교 및 유생으로부터 확인된 세균 규명)

  • Kwon, Sera;Park, Daesik;Choi, Woo-Jin;Park, Jae-Jin;Cho, Han-Na;Han, Ji-Ho;Lee, Jin-Gu;Koo, Kyo-Soung
    • Korean Journal of Environment and Ecology
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    • v.31 no.5
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    • pp.444-454
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    • 2017
  • There are many ongoing studies of infectious diseases as the major factor responsible for global declining of the amphibian population. Although some point out the amphibian rearing facilities like frog farms as one of the important sources of harboring and spreading amphibian infectious pathogens in the wild, there have been few related studies in South Korea. In this study, we investigated the bacterial and fungal colonies on the skin and in the internal organs of frogs and tadpoles collected inside and outside of Dybowski's brown frog farms in Inje, Goesan, and Gongju to compare the difference according to the region and between inside and outside the farm. We also intended to classify the bacteria collected from the tadpoles into species by analyzing 16s rDNA gene sequences. The result showed that the number of bacterial colonies found in the skin and gut of frogs and the number of fungal colonies found in the skin and liver of frogs collected in Goesan was significantly greater than those in the frogs in Inje. However, there was no difference between the frogs collected inside and outside of farms in both regions. In the case of tadpoles, the number of fungal colonies in the tadpoles collected from Gongju was greater than that in the tadpoles collected from Inje. The comparison of inside and outside frog farms showed that there were more bacterial colonies on the skin of the tadpoles collected from inside than outside the frog farm in Inje and more bacterial colonies in the organs of the tadpoles collected from outside than inside the farm in Gongju. The frogs with higher condition factor (body weight/snout-vent length*100) showed fewer bacterial colonies on the skin and fewer fungal colonies in the heart, but there were no significant relationships in tadpoles. We identified the total of 15 genera and four phyla of bacteria, but the difference according to regions and between inside and outside farm was not evident. The result of this study indicates that the different conditions according to the locality of farm and between inside and outside farm cause the difference in the population sizes of bacterial and fungal colonies and that it can affect the overall health condition of Dybowski's brown frogs in the farm. Moreover, the result suggests that effective disease control in the facility is greatly necessary to ensure successful operation of amphibian rearing facility and to prevent the possible spread of diseases from the facility to the wild.

Production of Violacein by a Novel Bacterium, Massilia sp. EP15224 Strain (Violacein을 생산하는 Massilia sp. EP15224 균주)

  • Yoon, Sang-Hong;Baek, Hee-Jin;Kwon, Soon-Wu;Lee, Chang-Muk;Sim, Joon-Soo;Hahn, Bum-Soo;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.317-323
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    • 2014
  • Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.

Conservation Scientific Diagnosis and Evaluation of Bird Track Sites from the Haman Formation at Yongsanri in Haman, Korea (함안 용산리 함안층 새발자국 화석산지의 보존과학적 진단 및 평가)

  • Lee, Gyu Hye;Park, Jun Hyoung;Lee, Chan Hee
    • Korean Journal of Heritage: History & Science
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    • v.52 no.3
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    • pp.74-93
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    • 2019
  • The Bird Track Site in the Haman Formation in Yongsanri (Natural Monument No. 222) was reported on the named Koreanaornis hamanensis and Jindongornipes kimi sauropod footprint Brontopodus and ichnospecies Ochlichnus formed by Nematoda. This site has outstanding academic value because it is where the second-highest number of bird tracks have been reported in the world. However, only 25% of the site remains after being designated a natural monument in 1969. This is due to artificial damage caused by worldwide fame and quarrying for flat stone used in Korean floor heating systems. The Haman Formation, including this fossil site, has lithofacies showing reddish-grey siltstone and black shale, alternately. The boundary of the two rocks is progressive, and sedimentary structures like ripple marks and sun cracks can clearly be found. This site was divided into seven formations according to sedimentary sequences and structures. The results of a nondestructive deterioration evaluation showed that chemical and biological damage rates were very low for all formations. Also, physical damage displayed low rates with 0.49% on exfoliation, 0.04% on blistering, 0.28% on break-out; however, the joint crack index was high, 6.20. Additionally, efflorescence was observed on outcrops at the backside and the northwestern side. Physical properties measured by an indirect ultrasonic analysis were found to be moderately weathered (MW). Above all, the southeastern side was much fresher, though some areas around the column of protection facility appeared more weathered. Furthermore, five kinds of discontinuity surface can be found at this site, with the bedding plane showing the higher share. There is the possibility of toppling failure occurring at this site but stable on plane and wedge failure by means of stereographic projection. We concluded that the overall level of deterioration and stability were relatively fine. However, continuous monitoring and conservation treatment and management should be performed as situations such as the physicochemical weathering of the fossil layer, and the efflorescence of the mortar adjoining the protection facility's column appear to be challenging to control.

A Study on the Setting Process and Formational Characteristics of the Seonyu Eight Scenic in Gogunsan Islands (고군산 선유팔경(仙遊八景)의 설정과정과 집경(集景) 특성)

  • Jung, Woo-Jin;Hwang, Guk-Woong
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.37 no.4
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    • pp.32-50
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    • 2019
  • The present study examines the circumstances around the establishment of the Seonyu Eight Scenic Spots (仙遊八景) in Gogunsan Islands and the characteristics of the landscape of each viewing point. The study conclusions are as follows. First, since the scenic spots were first established in 1969, their content and sequences have been changed several times, and their names have also been changed to some extent. Until the 1970s, these scenic spots did not have official names for them and were often specified as 'Gogunsan Eight Scenic Spots', and excluded 'the Musan Twelve Peaks (巫山十二峰)'. In addition, viewing points of the eight scenic spots varied across periods. This suggests that, for the early form of Seonyu Eight Scenic Spots, the picturesque scenery of Gogunsan Islands, and Seonyudo Island in particular, was chosen, while eight scenic spots in its vicinity were regarded. Second, the Seonyu Eight Scenic Spots of the early 2000s, which has all eight scenic spots of now, follows the nomenclature of the traditional eight scenic spots by specifying the sequence number with a refined name in four syllables. Its first scenic spot was Mangju Waterfall (望主瀑布) and its eighth scenic spot was Seonyu Sunset (仙遊落照); currently, the first scenic spot is Seonyu Sunset and the second scenic spot is Mangju Waterfall. Such change in the sequence of viewing points resulted from differences in representative landscape resources between the periods. Third, the lack of structure and finesse due to continuous changes is directed related to the identity issue of the Seonyu Eight Scenic Spots. Above all, it is unclear by whom and when Seonyu Eight Scenic Spots was established, and there are clear traces of following the eight scenic spots in the neighboring areas such as Okgu (沃溝) and Impi (臨陂)'s Eight Scenic Spots. Moreover, it is evaluated to have an unrefined, incomplete structure due to the lack of clarity in the knowledge and information about viewing objects, when to view, and historical and cultural background. Fourth, the first scenic spot, Seonyu Sunset, has the image that dominates the entire Eight Scenic Spots. The temporary landscape, the sunset, became the best view because it was perceived as the entirety of the landscape created by the fusion of the beautiful natural elements of Seonyudo Island. Therefore, there is ample room for raising the value of other landscape resources of Gogunsan Islands by utilizing the existing perception of exploring the entire landscape of Seonyudo Island and Gogunsan Islands starting with Seonyu Sunset. This likely requires additional work to imbue each viewing point with identity and completion.

Rapid Statistical Optimization of Cultural Conditions for Mass Production of Carboxymethylcellulase by a Newly Isolated Marine Bacterium, Bacillus velezensis A-68 from Rice Hulls (통계학적 방법을 사용한 해양미생물 Bacillus velezensis A-68균주의 섬유소 분해효소 생산 조건 최적화)

  • Kim, Bo-Kyung;Kim, Hye-Jin;Lee, Jin-Woo
    • Journal of Life Science
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    • v.23 no.6
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    • pp.757-769
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    • 2013
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater, identified as Bacillus velezensis by analyses of 16S rDNA and partial sequences of the gyrA, and designated as B. velezensis A-68. The optimal conditions for production of CMCase by B. velezensis A-68 were established using response surface methodology (RSM). The optimal concentrations of rice hulls and yeast extract, and initial pH of the medium for cell growth were 60.2 g/l, 7.38 g/l, and 7.18, respectively, whereas those for production of CMCase were 50.0 g/l, 5.00 g/l, and 7.30. The analysis of variance (ANOVA) implied that the most significant factor for cell growth as well as production of CMCase was yeast extract. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}7H_2O$, and $(NH_4)_2SO_4$ in the medium for cell growth were 7.50, 1.00, 0.10, and 0.80 g/l, respectively, which were the same as those for production of CMCase. The optimal temperatures for cell growth and production of CMCase were 30 and $35^{\circ}C$, respectively. The maximal production of CMCase under optimized conditions was 83.8 U/ml, which was 3.3 times higher than that before optimization. In this study, rice hulls, agro-byproduct, were developed as a substrate for production of CMCase and time for production of CMCase was reduced to 3 days using a newly isolated marine bacterium.

Analysis of the Amount of Telomeric DNA and Telomerase Activity on Preimplantation Mouse Embryoic Cells (마우스 수정란의 초기 배 발생단계별 Telomeric DNA의 양적 분석과 Telomerase 활성도 분석)

  • Kang M. Y.;Han M. S.;Lee S. C.;Kim J. H.;Sohn S. H.
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.1-7
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    • 2005
  • Telomeres consisting of (TTAGGG)n tandem repeat DNA sequences and associated proteins are essential for chromosome stability and related with cell senescence, apoptosis and cancer. The telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. This study was carried out to identify the distribution of telomeres on mouse chromosomes and also to analyze the amount of telomeres and telomerase activity of mouse embryos at early embryonic stages. Germ cells and early embryos from 1 cell to blastocyst stage were analyzed. The amount of telomeres was analyzed by quantitative fluorescence in situ hybridization technique(Q-FISH) using a human telomeric DNA probe, and telomerase activity was measured by telomeric repeat amplification protocol assay(TRAP). In results, the telomeres on mouse chromosomes were distributed at the ends of all autosomes and sex chromosomes. Although the quantity of telomeres varied among chromosomes, most of chromosomes had higher amount in q-arm telomeres than in p-arm telomeres. The results of Q-FISH indicated that the relative amount of telomeres of mouse embryos in each embryonic stage was approximately the same except the higher amount in blastocysts. Using TRAP assay on mouse embryos, telomerase activity was detected in all preimplantation stages from mature oocytes to blastocysts. Especially the telomerase activity was significantly increased at the morula and blastocyst stage. In conclusion, there may be a close association between the amount of telomeres and telomerase activity in early embryonic stages, and analysis of telomere quantity and telomerase activity on early development will be helpful for the investigation of embryogenesis and embryonic cell differentiation in mice.

Mouse Embryonic Stem Cell Uptakes of Buforin 2 and pEP-1 Conjugated with EGFP (생쥐 배아 줄기세포의 Buforin 2 및 pEP-1에 결합된 EGFP의 세포 내 수송)

  • Jung, Su-Hyun;Park, Seong-Soon;Lim, Hyun-Jung;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.11 no.2
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    • pp.111-119
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    • 2007
  • Differentiation of cells can be induced through modulation of endogenous regulators using exogenous factors. Useful transfection systems to transport a specific exogenous regulator into cell have been tried but still there are many obstacles to overcome. In this study, we examined the transfection efficiency of cell permeable peptides (CPPs) in mouse embryonic stem cell under the various conditions. To identify the CPP-mediated translocation of a protein, we employed recombinant CPP-enhanced green fluorescent protein (EGFP). Viability of R1 cells was different between experimental groups depending on the kind of CPP and the concentration of CPP-EGFP. Translocation of CPP-EGFPs into the R1 cells was not detected until 30 min after CPP-EGFPs treatment in all groups. After 1 hr, translocation of pEP-1-EGFP-N was detected, but it could not be detected in the other group. Transfection of pEP-1EGFP-N was independent on its concentration. The time course did not show saturation even after 24 hr in pEP-1-EGFP-N. These results showed that the permeability depended on the kind of CPP and the location of His-tag in the case of examined CPPs, and did not need biological energy. On summary, the efficiency of transfection of CPP-EGFP depends on the CPP sequences but the culture time is not a key factor in transfection for the mouse embryonic stem cell. For the future studies to improve the efficiency of translocation of protein into embryonic stem cells, it is needed to develop modified CPP or mediator. The studies would be very useful to induce the differentiation of embryonic stem cells.

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Development of Microsatellite Markers using BAC clone Sequencing on Porcine Chromosome 6q28 - 6q32 (돼지 6번 염색체(6q28 - 6q32)의 BAC clone 염기서열 분석에 의한 Microsatellite Markers 개발)

  • Chang, K.W.;Lee, K.T.;Park, E.W.;Choi, B.H.;Kim, T.H.;Cheong, I.C.;Oh, S.J.
    • Journal of Animal Science and Technology
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    • v.46 no.3
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    • pp.301-306
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    • 2004
  • This study was conducted to develop new markers at the region that was related to QTL affecting intramuscular fat and backfat thickness on chromosome 6q28 - 6q32 in pigs. Dozens of repeated sequences were founded using shotgun sequencing of several BAC clones corresponding to that region, of which five new microstellite markers that identified polymorphism were discovered. The mean number of alleles at each locus observed 2.13(KP0290F2), 4.63(KP0248Cll), 7.38(KP1231C91), 2.75(KPI23IC92) and 6.2S(KP1231C93) in 8 breeds(Landrace, Korean native pig, Duroc, Yorkshire, Berkshire, Wuzhishan pig, Xiang pig, Min pig). The average estimated heterozygosity values at each locus varied from 0.2100(KP0290F2) to 0.8304(KPI23IC91) in all populations. In other hand, the average allele of all loci WlL'I within range of 0.4517(Berkshire) and 0.6957 (Yorkshire). Of these markers, KP0248C11, KP1231C91 and KP1231C93 were identified to have optimal number of alleles, high heterozygosity values and low standard deviation values. Especially, KPI23IC91 and KPI231C93 might be considered as a useful marker for genetic mapping and diversity study.

Herpes Simplex Virus Thymidine Kinase Gene Therapy Delivered by Retroviral or Adenoviral Vector in Mouse Model of Lewis Lung Carcinoma (Lewis 폐암 마우스 모델에서 Retroviral Vector나 Adenoviral Vector로 이입된 Herpes Simplex Virus Thymidine Kinase 유전자치료)

  • Kwon, Hee-Chung;Jeong, Jae-Min;Kim, Jung-Hyeon;Ham, Yong-Ho;Seo, Ji-Sook;Lee, Ki-Ho;Kim, Chang-Min;Lee, Han-Soo;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.298-309
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    • 2000
  • Background : The antitumor effects of herpes simplex virus thymidine kinase (HSV-tk) and ganciclovir (GCV) strategies for cancer gene therapy have a the following advantages : 1) a direct cytotoxicity to HSV-tk modified cancer cells by GCV 2) a cell death by the local transfer of toxic metabolites from the HSV-tk modified cells to nearby unmodified tumor cells (bystander effect), and 3) in vivo bystander effect such as antitumor-immunity. Retroviral and adenoviral sequences can silence transgene expression in cells and mice. In this study, we investigated the above described advantages of HSV-tk/GCV strategy in Lewis lung cell and mouse lung cancer model using retroviral vector and adenoviral vector. Also, we observed whether the expression of a silenced gene can be reactivated by treating cells with butyrate. Methods : Retrovirus-HSV-tk and adenovirus-HSV-tk vectors were used for the transduction of Lewis lung carcinoma (LLC) cells. The change of HSV-tk expression by butyrate was measured by Western blol The antitumor activities containing bystander effect were observed in vivo (by MTT assay) and in vivo tumor models of various combinations of LLC and LLC-tk. Results : 1. Butyrate induced the enhancement of HSV-tk expression from adenovirally transduced cells but not from retrovirally transduced cells. 2. Both retrovirus-HSV-tk and adenovirus-HSV-tk vectors with GCV treatment were effective for killing of tumor cell in vitro and suppression of LLC tumorigenicity. Bystander effect was responsible for killing of mixture of LLC-tk and LLC in vitro and in vivo-tumorigenicity model. Conclusion : Butyrate could augment adenovirus-mediated HSV -tk gene expression. Cancer gene therapy with HSV-tk suicide gene by retroviral and adenoviral vector seems to be an effective approach for lung cancer therapy.

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