• Title/Summary/Keyword: 단백질 동정

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환경친화적 미생물 비료 개발을 위한 우모분해 세균의 분리 및 응용

  • U, Eun-Ok;Kim, Min-Ju;Yu, Eun-Yeon;Park, Geun-Tae;Son, Hong-Ju;Lee, Sang-Jun
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2007.05a
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    • pp.453-456
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    • 2007
  • 가금류의 도축 부산물로 대량 배출되고 있는 우모는 주로 생물학적으로 난분해성 단백질인 케라틴으로 구성되어 있다. 따라서 물리화학적 처리에 의하여 우모를 처리하고 있으나 이 방법은 환경오염을 유발한다. 따라서 본 연구에서는 환경친화적 우모 분해 공정을 개발하고, 우모 분해산물의 식물 성장을 위한 비료로서의 가치를 평가하기 위하여 퇴비화 볏짚에서 케라틴 분해효소 생성능이 우수한 균주인 RS7을 분리하였다. RS7의 생화학적 특성과 16S rDNA의 염기 서열을 분석하여 동정한 결과, Bacillus pumilus RS7로 동정되었다. B. pumilus RS7은 $30^{\circ}C$에서 배양 5일 만에 우모를 완전히 분해할 수 있었다. 본 균주에 의한 우모 분해산물은 Helianthus sannuus L.의 생장율, 잎 수 증가량, 개화율, 건조 생체량에 있어 대조군과 화학적 우모 분해산물보다 우수하였다. Bacillus pumilus RS7에 의한 우모 분해산물은 식물 성장 촉진을 위한 미생물기원 비료로써의 역할을 수행할 수 있었으며, 이에 따라 양계산업에서 배출되는 우모 폐기물이 환경에 주는 악영향을 감소시킬 수 있을 것으로 예측된다.

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Species Identification of Nontoxic Alexandrium tamarense (Dinophyceae) from Chinhae Bay, Korea, Using Molecular Probes (진해만에서 분리한 무독성 Alexandrium tamarense (Dinophyceae) 동정을 위한 molecular probe 이용)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.14 no.2
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    • pp.301-308
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    • 2004
  • The morphology of the apical pore complex, the first apical plate and the posterior sulcal plates in a new isolate of Alexandrium tamarense (Lebour) Balech from the Bay of Chinhae was compared with other that of toxic strains of A. tamarense previously isolated from Korean waters. Although this isolate was morphologically identical to these toxic strains, high performance liquid chromatography and mouse bioassay showed no evidence of toxin production. The nontoxic A. tamarense strain showed a strong positive binding activity with PNA lectin, indicating a high density of lactose and galactose residues on the cell surface, and in SDS-PAGE and Western blot analysis a unique protein of about 21-kDa molecular sizes was observed. These findings demonstrate that the use of PNA and immunobioassay could be used to discriminate between toxic and nontoxic strains of A. tamarense.

Biochemical and molecular characterization of a strain KA/S2 of Acnnthamoebc castellanii isolated from Korean soil (카스텔란니가시아메바(Acanthamoeba castellanii) 한국 토양분리주 KA/S2의 생화학적 및 분자생물학적 특성)

  • 정동일;공현희
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.79-86
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    • 1996
  • A strain, KA/S2, isolated from Korean soil and morphologically assigned to Acanthcmoebc cQsteLlcnii, was characterized by isoenzyme analysis , and total proteins profile, End mitochondrial (Mt) DNA restriction fragment length polymorphism (RFLP) , and compared with four reference strains assigned to the species (the authenitic Castellani, Neff, Ma, and Chang strains). It was found that four isoenzyme, total proteins, and Mt DNA RFLP patterns by eight restriction endonucleases of the strain KA/S2 were identical with those of the Neff strain, isolated from soil of California, USA. The Chang strain was unique in its morphology and total protein patterns. Interstrain polymorphisms of isoensyme profiles and Mt DNA RFLP patterns were observed among the Castellani, Neff, Ma, and Chang strains. Mt DNA RFLP was confirmed to be highly appropriate for the strain characterization and identification of Acnnthamoeba spry.

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Effect of Elevated Ultraviolet-B Radiation on Yield and Differential Expression of Proteome in Perilla (perilla frutescens L.) (잎들깨 수량과 단백질체 발현에 미치는 UV-B의 영향)

  • Hong, Seung-Chang;Hwang, Seon-Woong;Chang, An-Cheol;Shin, Pyung-Gyun;Jang, Byoung-Choon;Lee, Chul-Won
    • Korean Journal of Environmental Agriculture
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    • v.25 no.1
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    • pp.7-13
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    • 2006
  • Plastichouse cultivation for crops and vegetables in the winter has been widely popularized in Korea. In the vinylhouse Ultraviolet B penetration is lower than in the field, and so some problems, as plant overgrowth and outbreak of disease, occurred frequently. The effect of artificial supplement ultraviolet B $(UV-B:280{\sim}320nm)$ radiation on the physiological responses and yield of perilla (perilla frutescens) was investigated UV-B ray was radiated on perilla with the 10th leaf stage at the distance of 90, 120 and 150 cm from the plant canopy for 30 days after planting in the vinylhouse. The production of fresh perilla leaves was high in the order of plastic house, ambient+50% of supplemental UV-B, ambient ambient+100% of supplemental UV-B. Enhanced UV-B radiation affected the intensity of thirty-three proteins in 2-dimensional electrophoretic analysis of proteins and ten proteins out of them seemed to be responsive to UV-B : a protein was, ATP synthase CF1 alpha chain, down regulated and nine proteins (Chlorophyll a/b bindng protein type I, Chlorophyll a/b binding protein type II precursor, Photosystem I P700 chlorophyll a apoprotein A2, DNA recombination and repair protein recF, Galactinol synthase, S-adenosyl-L-methionine, Heat shock protein 21, Calcium-dependent protein kinase(CDPK)-like, Catalase) were up-regulated.

Cloning of Low-molecular-weight Glutenin Subunit Genes and Identification of their Protein Products in Common Wheat (Triticum aestivum L.) (보통 밀에서 저분자글루테닌 유전자 클로닝 및 단백질 동정)

  • Lee, Jong-Yeol;Kim, Yeong-Tae;Kim, Bo-Mi;Lee, Jung-Hye;Lim, Sun-Hyung;Ha, Sun-Hwa;Ahn, Sang-Nag;Nam, Myung-Hee;Kim, Young-Mi
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.547-554
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    • 2010
  • Low-molecular-weight glutenin subunit (LMW-GS) in common wheat (Triticum aestivum L.) is important for quality processing of bread and noodles. The objectives of this study were to clarify the composition of LMW-GSs and to identify their corresponding proteins. Using LMW-GS specific primers we cloned and characterized 43 LMW-GS genes in the wheat cultivar 'Jokyoung'. Some of these genes contain polypeptides different in size due to the presence of various deletions or insertions within repetitive and glutamine-rich domains. The comparison of deduced amino acid sequence of the LMW-GS genes in Jokyoung with that of 12 groups LMW-GSs of wheat cultivar Norin 61 showed that the deduced amino acid sequences were nearly the same to LMW-GS groups of 1, 2, 3/4, 5, 7, 10 and 11. All LMW-GS genes contain eight cysteine residues, which are conserved among all of the typical LMW-GS sequences. The relative positions of cysteine residues are also conserved, except those of the first and seventh. Based on phylogenetic analysis, the 43 sequences with the same N-terminal and C-terminal amino acid sequences were clustered in the same group. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 7 spots of LMW-GSs of Jokyoung separated by two-dimensional gel electrophoresis (2DE). Of them, Glu-B3 (LMW-m and LMW-s) and Glu-D3 (LMW-m) were detected in two and three spots, respectively and the others were not clear. Collectively, we classified diverse LMW-GSs and identified their corresponding protein products. These results will be helpful in breeding programs for improvement of wheat flour quality.

Proteome analysis of storage roots of two sweet potato cultivars with contrasting low temperature tolerance during storage (저온 저장 감수성 및 저항성 고구마 품종에서 저온 반응성 단백질체 연구)

  • Kim, Yun-Hee;Ji, Chang Yoon;Kim, Ho Soo;Chung, Jung-Sung;Choi, Sung Hwan;Kwak, Sang-Soo;Lee, Jeung Joo
    • Journal of Plant Biotechnology
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    • v.49 no.2
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    • pp.118-123
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    • 2022
  • To obtain information on the molecular mechanism underlying the low temperature tolerance of sweet potato [Ipomoea batatas (L.) Lam], the proteome expressed in the sweet potato cultivar Xushu 15-1 with high cold storage tolerance and in the cultivar Xushu 15-4 with low cold storage tolerance was analyzed using 2-D and MALDI-TOF/TOF analyses. Compared with the control (without cold treatment), four protein spots were newly expressed in Xushu 15-1. The expression level of one protein spot was higher in Xushu 15-4 than in Xushu 15-1. Spot 2, which was newly expressed in Xushu 15-1, was identified as sporamin. Assessment of the change in protein expression levels over 8 weeks in the storage roots of the two cultivars treated at 4℃ revealed no significant difference in the expression levels in Xushu 15-1 over time. However, in Xushu 15-4, the expression level of one protein spot increased, while those of four spots decreased. Of the proteins with reduced expression levels, spots 7 and 8 were identified as actin and spots 9 and 10 were identified as fructokinase-like proteins. The present results are expected to enhance the understanding of the complex mechanism underlying the low temperature tolerance of sweet potatoes during storage and can be used to identify candidate genes for the development of new varieties of sweet potatoes with improved low temperature tolerance during cold storage in the future.

Physiological and Proteomic Responses of Barley Seedlings to Salt Stress (보리의 생육초기 염 스트레스에 따른 생리적 반응과 프로테옴 변화)

  • Kim, Dea-Wook;Yun, Seong-Kun;Park, Hyoung-Ho;Hwang, Jong-Jin;Han, Ok-Kyu;Park, Tae-Il;Jung, Gun-Ho;Lee, Jae-Eun;Kim, Sun-Lim;Chung, Young-Ho
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.537-545
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    • 2011
  • This study was conducted to obtain basic information on physiological and proteomic responses of barley seedlings to salt stress. Shoot dry weight decreased significantly as the level of soil salinity increased. Salt stress-induced decrease of relative shoot dry weight was lower in cv. "Sanglok" than in cv. "Sunwoo". Under the salt stress, SPAD value decreased, and the value was higher in cv. "Sanglok" than in cv. "Sunwoo". Sodium ion content in the leaves increased as NaCl concentration increased, and the content was higher in cv. "Sunwoo" than in cv. "Sanglok". The K+/Na+ ratio was higher in cv. "Sanglok" than in cv. "Sunwoo". Salt stress-induced alterations in protein expression of the leaves were detected by two dimensional electrophoresis, and 47 protein spots showing altered expression were selected. Among the selected protein spots, 17 protein spots were up-regulated and 28 spots down-regulated in cv. "Sanglok". In cv. "Sunwoo", 14 protein spots were up-regulated and 27 spots down-regulated. Out of 47 deferentially expressed protein spots, 18 protein spots were identified using mass spectrometry and NCBI protein database. Among the identified proteins, ten proteins are known to be involved in various stress responses, but the others are not directly involved in stress responses.

Identification of Tumor Antigens in Lung Cancer Patient by SEREX (폐암 환자에서 면역항원유전자의 혈청학적 동정)

  • Min, Young-Ki;Ha, Jin-Mok;Son, Young-Ok;Park, Hae-Rim;Lee, Min-Ki;Park, Yeoung-Min;Kim, Cheol-Min;Lee, Sang-Yull
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1082-1089
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    • 2007
  • Serological anlysis of recombinant cDNA expression libraries (SEREX) has led to identification of several categories of new antigens recognized by the immune system of cancer patients, which are referred to as the cancer immunome. We analyzed normal testis cDNA expression libraries with serumobtained from non-small lung cancer patient and isolated 40 distinct antigen designated KP-LuT-1 through KP-LuT-40. Among these antigens 20 antigens were previously identified by SEREX analysis of other tumor types, and 20 out of 40 antigens (50%) did not match entries in Cancer Immunome Database and were considered newly identified antigens. Sequencing analysis showed that the anti-gens comprised 26 functional known proteins and 14 noble/uncharacterized gene products. Of these, the hypothetical protein KP-LuT-6 was shown tissue-restricted. RT-PCR showed it to be expressed strongly only in normal testis. In addition to normal tissues-restricted expression, KP-LuT-6 mRNA was detected in lung tumor samples(3/l0), stomach tumor samples(3/l0), and breast tumor samples(l/5), whereas not detected in colon tumor samples(O/I2). These data suggest that KP-LuT-6 is a cancer/testis (CT)-like antigen as a potential target for cancer immunotherapies.

Identification and characterization of laccase genes in the Flammulina velutipes var. lupinicola genome (Flammulina velutipes var. lupinicola의 유전체 정보기반 laccase 유전자 동정 및 특성 규명)

  • Yu, Hye-Won;Park, Young-Jin
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.285-293
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    • 2021
  • The purpose of this study was to identify and characterize the laccase genes of Flammulina velutipes var. lupinicola. Five laccase genes (g1934, g1937, g2415, g2539, g5858) were selected based on the copper binding site and signal peptide analysis results using the laccase gene selected from the F. velutipes var. lupinicola genome. The size of the laccase genes of F. velutipes var. lupinicola were 1,488 bp~1,662 bp. As a result of cDNA sequence analysis, 14 to 17 introns were identified in the laccase genes. The cleavage site predicted as the signal peptide of the laccase gene was found to be located between 20 bp and 34 bp from the N-terminus. In addition, separation and purification were performed to characterize the F. velutipes var. lupinicola laccases, and the optimal activity of the separated and purified proteins were analyzed by pH, temperature and time. Five bands with laccase activity were found from zymogram analysis. The optimal pH of the reaction was 5.5, the optimal temperature was found to be 40℃. Therefore, characterization of the laccase genes identified in this study should help in better understanding the biomass decomposition of F. velutipes var. lupinicola.

Nucleotide Sequence of an Extracellular Phospholipase D Gene from Streptomyces somaliensis and Transphosphatidylation Activity of Its Enzyme (Streptomyces somaliensis가 생산하는 세포외 Phospholipase D의 유전자 서열 분석과 Transphosphatidylation 활성 특성)

  • Jeong Sujin;Lee Sun-Hee;Uhm Tai-Boong
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.211-216
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    • 2004
  • A bacterial strain JE-ll found to produce active extracellular phospholipase D (PLD) was selected from the soil isolates. It was identified as Streptomyces somaliensis on the basis of 16S rDNA sequence analysis, morphological and physiological characteristics. The gene (sspld) encoding S. somaliensis PLD was isolated and characterized. The open reading frame was suggested to encode 538 amino acids with a signal peptide of 33 amino acids. The deduced amino acid sequence of the sspld shared a sequence similarity of 70-88% with PLDs of other Streptomyces sp. so far reported. The PLD converted phosphatidylcholine to phosphatidylglycerol or phosphatidylserine with the yield of 96 to 99% (㏖/㏖), but did not act on inositol or ethanolamine as a transphosphatidylation donor.