• Title/Summary/Keyword: 단백질 동정

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Characterization and Condition of Silver Accumulation Bacteria in Groundwater (지하수에서 분리한 은(銀) 축적균주에 대한 축적조건 및 특성)

  • 배진희;민병례;한명수;최영길
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.273-276
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    • 2001
  • The strain which accumulate the silver in cell were isolated and characterized. And condition of accumulation of heavy metal was examined closely to investigate optimal condition of accumulation. Pseudomonas fluorescens and Bacillus cereus were Isolated from groundwater as the strain of silver accumulating bacteria. These strains did not grow in the medium at silver over the concentration 20 ppm. The largest accumulations of silver in the culture of Pseudomonas fluorescens and Bacillus cereus occurred within 24 hours. The amount of silver accumulation in Pseudomonas fluorescens and Bacillus cereus were 1.9 mg/g cell and 1.65 mg/g cell, repletively. In protein patterns of cell after the treatment of silver, three reducible proteins (126 KDa, 89 KDa, 25 KDa) in Bacillus cereus and one new protein (34 KDa) in Pseudomonas fluorescens were detected by SDS-PAGE.

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Identification of a Potential Tyrosine Phosphorylation Site on the NR2B Subunit of the N-methyl-D-aspartate Receptor (NMDA 수용체 아단위 2B의 Tyrosine 인산화 위치의 동정)

  • Il Soo Moon;Yong Wook Jung;Bok Hyun Ko
    • Journal of Life Science
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    • v.8 no.6
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    • pp.654-659
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    • 1998
  • The 2B subunit of N-methyl-D-aspartate (NMDA) receptors (NR2B) is the major phosphotyrosine-containing pro-tein in the postsynaptic density (PSD). In order to identify the site for tyrosine phosphorylation on NR2B, a mass spectrometry was applied on tryptic and endolys-C peptides. The NR2B subunit was isolated from N-octyl glucoside (NOG)-insoluble PSD fraction through SDS-PAGE and electroelution. The eluted protein was confirmed to be NR2B and phosphorylated on tyrosine by its cognate antibody and phosphotyrosine-specific antibody. By matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry of the peptides generated by digesting the eluted NR2B with trysin or endolys-C, a potential site for tyrosine phosphorylation could be identified as Tyr-1304.

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groES Expression Related to Antifungal Activity of Streptomyces sp. SAR01 (Streptomyces sp. SAR01 균주에서의 항진균 관련 groES의 발현)

  • 이영근;김재성;조규성;장병일;추철형
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.162-167
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    • 2002
  • To analyse proteins and gene related to antifungal activity, SAR01 strain was isolated from a brown seaweed and identified as Streptomyces sp. by FAME(fatty acid methyl ester) analysis. Antifungal activity deficient mutant(SAR535) of Streptomyces sp. SAR01 was induced by gamma radiation$({60}^Co)$. It was found that 6 specific protein spots appeared only in SAR01 by 2-D electrophoresis analysis. Among them, a protein of 10 kDa had homology of 96% with 10 kD chaperonin cpn 10 (GroES) by Basic Local Alignment Search Tool(BLAST, NCBI) analysis. SAR535 transformants into which groES was transferred by electroporation revealed antifungal activity newly similar with SAR01 It suggested that groES be supposed to be related to the antifungal activity of Streptomyces sp. SAR01.

Proteome Characterization of Sorghum (Sorghum bicolor L.) at Vegetative Stage under Waterlogging Stress (토양 과습 조건하에서 수수 잎의 단백질 양상)

  • Yun, Min Heon;Jeong, Hae-Ryong;Yoo, Jang-Hwan;Roy, Swapan Kumar;Kwon, Soo-Jeong;Kim, Joo-Ho;Chun, Hyen Chung;Jung, Ki Yuol;Cho, Seong-Woo;Woo, Sun-Hee
    • Korean Journal of Plant Resources
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    • v.31 no.2
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    • pp.124-135
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    • 2018
  • The study was performed to explore the molecular changes in the vegetative stage (3-and 5-leaf) of sorghum under waterlogging stress. A total of 74 differentially expressed protein spots were analyzed using LTQ-FT-ICR MS. Among them, 12 proteins were up-regulated and 3 proteins were down-regulated. Mass spectrometry (MS) results showed that about 50% of the proteins involved in various metabolic processes. The level of protein expression of malate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase related to carbohydrate metabolic process increased in both 3 and 5-leaf stage under waterlogging stress. These proteins are known to function as antistress agents against waterlogging stress. The expression of oxygen-evolving enhancer protein 1 protein related to photosynthesis was slightly increased in the treated group than in the control group, however the expression level was increased in the 5-leaf stage compared to the 3-leaf stage. Probable phospholipid hydroperoxide glutathione peroxidase protein and superoxide dismutase protein related to response to oxidative stress showed the highest expression level in 5-leaf stage treatment. This suggests that the production of reactive oxygen species by the waterlogging stress was the most abundant in the 5-leaf treatment group, and the expression of the antioxidant defense protein was increased.

Analysis of Antifungal Proteins in Streptomyces sp. SAR01 (Streptomyces sp. SAR01 균주에서의 항진균 관련 단백질 분석)

  • 이영근;김재성;조규성;장병일;추철형
    • Korean Journal of Environmental Biology
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    • v.20 no.3
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    • pp.237-244
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    • 2002
  • To analyze proteins related to antifungal activity, SAR01 strain was isolated from seaweed and identified as Streptomyces sp. from the result of FAME (fatty acid methyl ester) analysis. The isolated strain had antifungal activities against T species of plant pathogenic fungi. Antifungal activity deficient mutant (SAR 535) of Streptomyces sp. SAR01 was induced by gamma radiation $(^{60}Co,\;5kGy)$. By 2 D electrophoresis analysis, 6 protein spots were found in wild strain (SAR01) but these spots disappeared in mutant strain (SAR535). Among them, 5 proteins showed similarities to heat shock protein 70(HSP70), Fe-containing superoxide dismutase II (Fe- SODII), ribosome recycling factor (RRF), 10 kDa chnperonin (GroES) and inorganic pyrophosphatase (PPAse), respectively. It suggested that the above 6 proteins could be closely related to the antifungal activity of Streptomyces sp. SAR01.

Identification of Upregulated APOA1 Protein of Chicken Liver in Pullorum Disease (추백리가 감염된 닭의 간에서 발현이 증가하는 APOA1 단백질의 확인)

  • Jung K. C.;Lee Y. J.;Yu S. L.;Lee J. H.;Jang B. K.;Koo Y. B.;So H. K.;Choi K. D.
    • Korean Journal of Poultry Science
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    • v.32 no.1
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    • pp.23-27
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    • 2005
  • The aim of this study was to investigate differentially expressed proteins between normal chicken liver and chicken liver inffeted by Salmonella pullorum. 2-dimensional electrophoresis (2DE) and mass spectrometry (MS) were used to identify the proteins. More than 300 protein spots were detected on silver stained 2DE gels using pH 3$\~$10 gradients. The most outstanding protein spot was further analyzed by MALDI-TOF MS and protein database using the Mascot search engine. The protein was finally identified as APOAI (Apolipoprotein AI). Based on the known function of the APOAI, this gene acts protective action against the accumulation of platelet thrombin at the site of vascular damage for the pullorum disease. Therefore APOAI protein, identified in this study, can be a valuable biomarker in relation to the pullorum disease in chicken.

The enhancement of protein separation by duplex SDS-PAGE (Duplex SDS-PAGE를 이용한 단백질 분리향상)

  • Pyo, Jae Sung;Roh, Si Hun;Song, Jin-Su;Lee, Kyung Hyeon;Kim, Hie-Joon;Park, Jeong Hill;Kwon, Sung Won
    • Analytical Science and Technology
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    • v.19 no.6
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    • pp.529-534
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    • 2006
  • The protein separation with molecular weight using SDS-PAGE(sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is the one of the most conventional and simple techniques. In, this study, two dimensional SDS-PAGE using same separation principle consecutively was investigated and compared with one dimensional SDS-PAGE. The enhanced separation from duplex SDS-PAGE was observed and separated proteins in the gel were identified by MALDI TOF MS. Identified proteins from different gel spots were found to have different gi numbers. Therefore, duplex SDS-PAGE separation method will be used for economic separation method in the future because only tiny amount of inexpensive reagents are used to perform duplex SDS-PAGE.

Identification and characteristics of DDX3 gene in the earthworm, Perionyx excavatus (팔딱이 지렁이(Perionyx excavatus) DDX3 유전자의 동정 및 특성)

  • Park, Sang Gil;Bae, Yoon-Hwan;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.23 no.1
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    • pp.70-81
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    • 2015
  • Helicases are known to be a proteins that use the chemical energy of NTP binding and hydrolyze to separate the complementary strands of double-stranded nucleic acids to single-stranded nucleic acids. They participate in various cellular metabolism in many organisms. DEAD-box proteins are ATP-dependent RNA helicase that participate in all biochemical steps involving RNA. DEAD-box3 (DDX3) gene is belonging to the DEAD-box family and plays an important role in germ cell development in many organisms including not only vertebrate, but also invertebrate during asexual and sexual reproduction and participates in stem cell differentiation during regeneration. In this study, in order to identify and characterize DDX3 gene in the earthworm, Perionyx excavatus having a powerful regeneration capacity, total RNA was isolated from adult head containing clitellum. Full length of DDX3 gene from P. excavatus, Pe-DDX3, was identified by RT-PCR using the total RNA from head as a template. Pe-DDX3 encoded a putative protein of 607 amino acids and it also has the nine conserved motifs of DEAD-box family, which is characteristic of DEAD-box protein family. It was confirmed that Pe-DDX3 has the nine conserved motifs by the comparison of entire amino acids sequence of Pe-DDX3 with other species of different taxa. Phylogenetic analysis revealed that Pe-DDX3 belongs to a DDX3 (PL10) subgroup of DEAD-box protein family. And it displayed a high homology with PL10a, b from P. dumerilii.

Isolation of the Protease-producing Yeast Pichia anomala CO-1 and Characterization of Its Extracellular Neutral Protease (세포 외 중성 단백질분해효소를 생산하는 Pichia anomala CO-1의 분리 동정 및 효소 특성)

  • Kim, Ji Yeon
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1126-1135
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    • 2019
  • From a sample of bamboo byproduct, the protease-producing yeast strain CO-1 was newly isolated. Strain CO-1 is spherical to ovoid in shape and measures $3.1-4.0{\times}3.8-4.4{\mu}m$. For the growth of strain CO-1, the optimal temperature and initial pH were $30^{\circ}C$ and 4.0, respectively. The strain was able to grow in 0.0-15.0%(w/v) NaCl and 0.0-9.0%(v/v) ethanol. Based on a phylogenetic analysis of its 18S rDNA sequences, strain CO-1 was identified as Pichia anomala. The extracellular protease produced by P. anomala CO-1 was partially purified by ammonium sulfate precipitation, which resulted in a 14.6-fold purification and a yield of 7.2%. The molecular mass of the protease was recorded as approximately 30 kDa via zymogram. The protease activity reached its maximum when 1.0%(w/v) CMC was used as the carbon source, 1.0%(w/v) yeast extract was used as the nitrogen source, and 0.3%(w/v) $MnSO_4$ was used as the mineral source. The protease revealed the highest activity at pH 7.0 and $30^{\circ}C$. This enzyme maintained more than 75% of its stability at a pH range of 4.0-10.0. After heating at $65^{\circ}C$ for 1 hr, the neutral protease registered at 60% of its original activity. The protease production coincided with growth and attained a maximal level during the post-exponential phase.

in vitro 에서 Human lactadherin gene의 발현 및 동정

  • 전길수;염행철
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.93-93
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    • 2003
  • 모유의 뮤신 복합체는 rotavirus에 특이적으로 결합하여 항 바이러스활동을 보여주는 것으로 나타났다. 자연 상태에서의 뮤신은 몇몇 작은 분자들과 복합적으로 연합되어 있는데 70kda의 glycoprotein, butyrophilin, 그리고 glycosylated component, lactadherin을 포함하고 있다. 그중 rotavirus에 가장 높은 결합력과 항바이러스 활동을 나타내는 Lactadherin은 모유의 유단백질의 하나인 뮤신과 결합되어 분비되는 당단백질의 하나로 분자량이 46kda이고, 지방구막 속에 연합되어있다. 이러한 배경에서 본 연구에서는 한국 여성의 breast tissue로부터 lactadherin 유전자의 cloning 및 in vitro에서의 발현을 유도하여 lactadherin band만을 purify하였고 여기서 얻은 lactadherin을 항체 생산을 위한 항원으로 사용하여 anti-lactadherin antibody를 확보하였다. 이 항체는 human milk에서의 lactadherin을 동정하는데 사용하였는데 western blot결과 lactadherin을 포함하여 몇 몇 단백질들이 확인되었다. Human milk내 mucin은 몇몇 작은 분자들과 복합체를 형성하는 것으로 확인되어졌는데 70kda의 glycoprotein, butyrophilin 그리고 46kda의 glyxosylated component, lactadherin을 포함하고 있는 milk mucin은 associated molecular임을 확인할 수 있었다.

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