• 제목/요약/키워드: Isolated pore

검색결과 64건 처리시간 0.031초

Cloning of a pore-forming subunit of ATP-sensitive potassium channel from Clonorchis sinensis

  • Hwang, Seung-Young;Han, Hye-Jin;Kim, So-Hee;Park, Sae-Gwang;Seog, Dae-Hyun;Kim, Na-Ri;Han, Jin;Chung, Joon-Yong;Kho, Weon-Gyu
    • Parasites, Hosts and Diseases
    • /
    • 제41권2호
    • /
    • pp.129-133
    • /
    • 2003
  • A complete cDNA sequence encoding a pore-forming subunit (Kir6.2) of ATP-senstive potassium channel in the adult worm, Clonorchis sinensis, termed CsKir6.2, was isolated from an adult CDNA library. The cDNA contained a single open-reading frame of 333 amino acids, which has a structural motif (a GFG-motif) of the putative pore-forming loop of the Kir6.2. Peculiarly, the Cskir6.2 shows a lack-sequence structure, which deleted 57 amino acids were deleted from its N-terminus. The predicted amino acid sequence revealed a highly conserved sequence as other known other Kir6.2 subunits. The mRNA was weekly expressed in the adult worm.

Purification of a Pore-forming Peptide Toxin, Tolaasin, Produced by Pseudomonas tolaasii 6264

  • Cho, Kwang-Hyun;Kim, Sung-Tae;Kim, Young-Kee
    • BMB Reports
    • /
    • 제40권1호
    • /
    • pp.113-118
    • /
    • 2007
  • Tolaasin, a pore-forming peptide toxin, is produced by Pseudomonas tolaasii and causes brown blotch disease of the cultivated mushrooms. P. tolaasii 6264 was isolated from the oyster mushroom damaged by the disease in Korean. In order to isolate tolaasin molecules, the supernatant of bacterial culture was harvested at the stationary phase of growth. Tolaasin was prepared by ammonium sulfate precipitation and three steps of chromatograpies, including a gel permeation and two ion exchange chromatographies. Specific hemolytic activity of tolaasin was increased from 1.7 to 162.0 HU $mg^{-1}$ protein, a 98-fold increase, and the purification yield was 16.3%. Tolaasin preparation obtained at each purification step was analyzed by HPLC and SDS-PAGE. Two major peptides were detected from all chromatographic preparations. Their molecular masses were analyzed by MALDI-TOF mass spectrometry and they were identified as tolaasin I and tolaasin II. These results demonstrate that the method used in this study is simple, time-saving, and successful for the preparation of tolaasin.

New record of the cold freshwater dinoflagellate Palatinus apiculatus (Dinophyceae) from the Paldang Reservoir, Korea

  • Kim, Taehee;Ki, Jang-Seu
    • Journal of Species Research
    • /
    • 제11권3호
    • /
    • pp.162-168
    • /
    • 2022
  • Compared to marine dinoflagellates, freshwater species are rarely recorded in Korea. In the present study, we isolated a freshwater dinoflagellate, Palatinus, from the Paldang Reservoir, Korea, in December 2021. The overall cell shape was ovoid, and the cell size was 34.3 ㎛ in length (25.8-39.5 ㎛) and 28.4 ㎛ in width (21.5-34 ㎛). An eyespot was usually observed near the sulcal region. The Kofoidian plate formula of the species was determined to be 4', 2a, 7", 6c, 5s, 5''', and 2''''. Apical pore complex was not observed. However, variations in the cingular plate caused by the fusion of 3C and 4C were observed. Analyses of 28S rDNA sequences revealed that the unidentified species is 100% similar to Palatinus apiculatus, and clustered together in the same lineage in the phylogenetic tree (100% bootstrap value). Our findings confirmed that the isolated dinoflagellate is Palatinus apiculatus, which was discovered for the first time in Korean freshwaters.

Ultrastructure of the Integument of Capillaria hepatica (syn. Calodium hepatica) (간모세선충(Capillaria hepatica) 표피의 미세구조)

  • Kim, Soo-Jin;Min, Byoung-Hoon;Lee, Haeng-Sook;Lee, Byoung-Wook;Joo, Kyoung-Hwan
    • Applied Microscopy
    • /
    • 제39권2호
    • /
    • pp.167-173
    • /
    • 2009
  • Capillaria hepatica is a parasitic nematode which causes hepatic capillariasis in rodents and other mammals, including man. Rat species of the genus Rattus are main primary host and rates of genus Rattus of up to 100% have been reported. Infection to reservoir and other mammalian hosts occur incidentally due to ingestion of water or food contaminated with C. hepatica embryonated eggs. The worms mature exclusively inside the liver, but they die and disassemble soon after egg spawning in rats. Dead worms and their eggs cause immune response of focal necrosis and inflammation within the liver. C. hepatica adult with a thin and long body is similar to capillary. The members of Order Trichurida are characterized by having a stichosome and the bacillary bands in front of the body. As already mentioned, the adult C. hepatica residesin the liver, where it deposits groups of eggs, and finally die in the encapsulated tissue of the liver. They produce eggs that elicit a marked granulomatous reaction that eventually destroy the worms. And the adult worms were mixed with eggs. So the complete isolation of the worm and observation of intact ultrastructure is very difficult. In this study, integument structure of C. hepatica isolated from the liver of mouse at 7 weeks after inoculation of embryonated eggs were observed with scanning and transmission electron microscopy. As a results, body length of isolated C. hepatica was about 99 mm. Cuticle, bacillary band and bacillary pore were obtained in the integument of worm. Bacillary pore across cuticular surface of the worm were observed. According to the existence of cap material, external forms of bacillary pore can be divided into three types such as flat, ingression, and ingression with the cap material type. The complete isolation of the worm and observation of ultrastructure of integument will provide the fundamental data which is important in the nematode research including C. hepatica.

Property of MgO with Different Sintering Temperatures under High Pressures (고압 환경에서 소결 온도에 따른 MgO 물성의 변화)

  • Song, Jeongho;Noh, Yunyoung;Song, Ohsung
    • Journal of the Korean Ceramic Society
    • /
    • 제49권6호
    • /
    • pp.608-613
    • /
    • 2012
  • We investigated the property changes of MgO powders sintered at temperatures ranging from $700^{\circ}C$ to $1900^{\circ}C$ for 5minutes at a pressure of 2.7 GPa for a high-pressure high-temperature(HPHT) diamond synthesis process. The physical properties of the sintered MgO powders were characterized by optical microscopy, field emission scanning electron microscopy (FE-SEM), Vickers hardness tests, and by the apparent density, and X-ray diffractometry. An optical micro-analysis showed that white MgO powders became black after sintering due to carbon contamination from the graphite heat source. FE-SEM revealed the growth in the grain size of the MgO powders from $0.3{\mu}m$ to $50{\mu}m$ after sintering at $1700^{\circ}C$. The hardness and apparent density increased to $1800^{\circ}C$ while the samples were dedensified at $1900^{\circ}C$ due to the growth of isolated pores. According to the XRD analysis, no phase transformation occurred in the MgO powders. These results suggest that HPHT-sintered MgO powders can show an accelerated sintering process characterized by grain neck growth, pore connections, isolated pore growth and dedensification in 5 minutes, while these processes with the conventional sintering process take at least 5 hours.

The Production of Functional Peptide from Whey Using Immobilized Trypsin (유청으로부터 고정화 트립신을 이용한 기능성 펩타이드의 생산)

  • Park, Yun-Joo;Yun, Yeo-Pyo;Lee, Hyung-Joo;Jang, Hae-Dong
    • Korean Journal of Food Science and Technology
    • /
    • 제28권1호
    • /
    • pp.99-104
    • /
    • 1996
  • Carbohydrate-free caseinomacropeptide (CMP) was isolated from the sweet whey powder by a precipitation method using 12% trichloroacetic acid. The yield of carbohydrate-free CMP was 2.7 g from 100 g sweet whey powder. The electrophoretic pattern and the amino acid analysis of CMP showed that isolated CMP was quite pure, indicating the precipitation with 12% trichloroacetic acid was very effective for isolating carbohydrate-free CMP from the sweet whey powder. Trypsin, covalently immobilized on pore glass beads by carbodiimide (EDC) method, was 20mg per 1g glass beads. CMP was almost completely hydrolyzed by soluble trypsin in 24hr, but not by immobilized trypsin. The tryptic hydrolysates were fractionated on a Bio-Gel P 4 column $(1.5{\times}120\;cm)$and separated peptides were tested for their capacities to inhibit platelet aggregation using a aggregometer. The hydrolysate obtained from CMP after 24hr digestion by immobilized trypsin showed the highest activity.

  • PDF

A STUDY ON NUMERICAL COUPLING BETWEEN MECHANICAL AND HYDRAULIC BEHAVIORS IN A GRANITE ROCK MASS SUBJECT TO HIGH-PRESSURE INJECTION

  • Jeong, Woo-Chang;Jai-Woo;Song, Jai-Woo
    • Water Engineering Research
    • /
    • 제2권2호
    • /
    • pp.123-138
    • /
    • 2001
  • An injection experiment was carried ut to investigate the pressure domain within which hydromechanical coupling influences considerably the hydrologic behavior of a granite rock mass. The resulting database is used for testing a numerical model dedicated to the analysis of such hydromechanical interactions. These measurements were performed in an open hole section, isolated from shallower zones by a packer set at a depth of 275 m and extending down to 840 m. They consisted in a series of flow meter injection tests, at increasing injection rates. Field results showed that conductive fractures from a dynamic and interdependent network, that individual fracture zones could not be adequately modeled as independent systems, that new fluid intakes zones appeared when pore pressure exceeded the minimum principal stress magnitude in that well, and that pore pressures much larger than this minimum stress could be further supported by the circulated fractures. These characteristics give rise to the question of the influence of the morphology of the natural fracture network in a rock mass under anisotropic stress conditions on the effects of hydromechanical couplings.

  • PDF

Filter Plate Micro Trap as a Device for in situ Cultivation for Environmental Microorganisms (환경시료에 존재하는 미생물 배양을 위한 filter plate micro trap의 개발)

  • Jung, Da-Woon;Ahn, Tae-Seok
    • Journal of Life Science
    • /
    • 제22권6호
    • /
    • pp.723-729
    • /
    • 2012
  • Filter plate microbial trap (FPMT) was invented as an in situ cultivation device for the isolation of bacteria from natural environments. FPMT consists of a medium and membrane filters (0.45 ${\mu}m$ pore size) and microorganisms and compounds can be moved freely moved into the medium. This device was applied to two soil samples of Greenland. The microbial diversity of both soil samples by FPMT was higher than that by the conventional Petri dish-based method. Moreover, novel bacterial species were isolated by FPMT. The new FPMT is effective for in situ cultivation of natural samples and could be applicable to the isolation of uncultivable microorganism.

Various Pathogenic Pseudomonas Strains that Cause Brown Blotch Disease in Cultivated Mushrooms

  • Mu, Lin-Lin;Yun, Yeong-Bae;Park, Soo-Jin;Cha, Jae-Soon;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
    • /
    • 제58권4호
    • /
    • pp.349-354
    • /
    • 2015
  • Brown blotch disease in cultivated mushrooms is caused by Pseudomonas tolaasii, which secretes a lipodepsipeptide, tolaasin. Tolaasin is a pore-forming toxin in the cell membranes, thus destroying the fruiting body structure of mushroom. In this study, we isolated pathogenic bacteria from mushrooms that had symptoms of brown blotch disease. In order to identify these bacteria, their 16S rRNA genes were sequenced and analyzed. Pathogenic bacteria identified as Pseudomonas species were thirty five and classified into five subgroups: P1 to P5. Each subgroup showed different metabolic profile measured by API 20NE kit. Fifty percent of the bacteria were identified as P. tolaasii (P1 subgroup). All five subgroups caused the formation of brown blotches on mushroom tissues and the optimum temperature was 25oC, indicating that they may be able to secrete causal factors, such as tolaasin and similar peptide toxins. These results show that there are at least five different pathogenic Pseudomonas species as blotch-causing bacteria and, therefore, strains from the P2 to P5 subgroups should be also considered and studied as pathogens in order to improve the quality and yield of mushroom production.

A Novel Antifungal Analog Peptide Derived from Protaetiamycine

  • Lee, Juneyoung;Hong, Hyun Joo;Kim, Jin-Kyoung;Hwang, Jae-Sam;Kim, Yangmee;Lee, Dong Gun
    • Molecules and Cells
    • /
    • 제28권5호
    • /
    • pp.473-477
    • /
    • 2009
  • Previously, the 9-mer analog peptides, 9Pbw2 and 9Pbw4, were designed based on a defensin-like peptide, protaetiamycine isolated from Protaetia brevitarsis. In this study, antifungal effects of the analog peptides were investigated. The antifungal susceptibility testing exhibited that 9Pbw4 contained more potent antifungal activities than 9Pbw2. A PI influx assay confirmed the effects of the analog peptides and demonstrated that the peptides exerted their activity by a membrane-active mechanism, in an energy-independent manner. As the noteworthy potency of 9Pbw4, the mechanism(s) of 9Pbw4 were further investigated. The membrane studies, using rhodamine-labeled giant unilamellar vesicle (GUV) and fluorescein isothiocyanate (FITC)-dextran loaded liposome, suggested that the membrane-active mechanism of 9Pbw4 could have originated from the pore-forming action and the radii of pores was presumed to be anywhere from 1.8 nm to 3.3 nm. These results were confirmed by 3D-flow cytometric contour-plot analysis. The present study suggests a potential of 9Pbw4 as a novel antifungal peptide.