• Title/Summary/Keyword: 단백질체

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Anti-Varicella Zoster Virus Activity of Water Soluble Components of Elfvingia applanata Alone and in Combinations with Interferons (잔나비걸상 수용성성분의 항-Varicella Zoster Virus 작용과 Interferon과의 병용효과)

  • Kim, Young-So;Lee, Seong-Kug;Lee, Young-Nam;Han, Seong-Sun
    • The Korean Journal of Mycology
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    • v.27 no.3 s.90
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    • pp.237-241
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    • 1999
  • To search for less toxic antiviral agents from Basidiomycetes, the water soluble components (=EA), were isolated from the carpophores of Elfvingia applanata (Pers.) Karst. Anti-varicella zoster virus (Oka strain; anti-VZV/Oka) activity of EA was examined in MRC-5 cells by plaque reduction assay in vitro. And the combined antiviral effects of EA with interferon (IFN) alpha or IFN gamma were examined on the multiplication of VZV/Oka. EA exhibited a dose-dependent reduction in the plaque formation of VZV/Oka with 50% effective concentration $(EC_{50})$ of $464.14\;{\mu}g/ml$. The results of the combination assay were evaluated by the combination index (CI) that was calculated by the multiple drug effect analysis. The combination of EA with IFN alpha showed partially synergistic or additive effects with CI values of $0.83{\sim}1.09$ for 50%, 70%, 90% effective levels, and those with IFN gamma showed antagonism with CI values of $1.20{\sim}1.24$.

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Purification and Characterization of Superoxide Dismutase in Sphingomonas sp. KS 301 (Sphingomonas sp. KS 301의 Superoxide Dismutase 정제 및 특성)

  • Kang, Hee-Jeong;Jeong, Jae-Hoon;Choi, Ji-Hye;Son, Seung-Yeol
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.83-90
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    • 2007
  • Sphingomonas sp. KS 301, which was isolated from oil contaminated soil, was shown to have five different SODs (SODI, II, III, IV, V) which can be separated by DEAE-Sepharose chromatography, and SOD III was finally purified in this study by ammonium sulfate precipitation, DEAE-Sepharose chromatography, Superose 12 gel filtration and Uno-Q1 ion exchange chromatography. The molecular weight of SOD III was 23 kDa as determined by SDS-PAGE and the apparent molecular weight of the native enzyme was estimated to be approximately 71 kDa by Superose-12 gel filtration chromatography. These data suggest that the purified SOD consists of at least two subunits. The specific activity of the SOD III was higher than Mn type or Fe type SOD of Escherichia coli by 5 fold. To determine the type of SOD III, inhibitory effects of $NaN_{3},\;H_{2}O_{2},\;KCN$ were examined. 10 mM $NaN_{3}$ was able to inhibit 56% of the SOD III activity, which indicates that this SOD is Mn type. The optimum pH of the SOD III was 7.0 and the optimum temperature was $20^{\circ}C$. N-terminal amino acid sequence of purified SOD III was most similar to those of Psudomonase ovalis and Vibrio cholerae among bacteria.

Current Research Status on the Development of Genetically Modified Plants in Korea (유전자변형식물의 국내 연구 현황)

  • Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.1-6
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    • 2003
  • In an attempt to evaluate the current research status of genetically modified (GM) plants, the scientific research publications in Korea as well as in international SCI journals were screened. About 190 research articles related to the development of GM plants were searched from 10 different domestic journals in the last 12 years (Jan. 1990 to Sept. 2002), The researches in 65 articles were carried out with tobacco plant, 20 with rice, 19 with potatoes, and less then 9 articles from each other plant species, respectively, In total, 38 different plant species were being subjected for the development of GM plants. In particular, there was only one article for each major staple grains such as wheat, barley, soybean, and maize. In more than 47% of total published articles, scientists mainly focused on the basic research such as developing transformation system (46 articles), gene expression study in transgenic plants (34), and vector constructions (10). In addition, 28 articles which main authors are Korean scientists were searched from 11 different international SCI journals. Again, major plants for GM research were tobacco (10) and rice (7). More than 50% of published articles were focused on the basic research, gene expression study with transgenic plants (16). The publications on the research of disease-resistant plants were 7 articles, 3 for the development of stress-resistant and 2 for the herbicide-resistant plants, respectively. It is believed that the last 10 year's investment through government organizations has just strengthen the capacity for the next big stride on agricultural biotechnology in Korea.

AtCBP63, a Arabidopsis Calmodulin-binding Protein 63, Enhances Disease Resistance Against Soft Rot Disease in Potato (애기장대 칼모듈린 결합 단백질 AtCBP63을 발현시킨 형질전환 감자의 무름병 저항성 증가)

  • Chun, Hyun-Jin;Park, Hyeong-Cheol;Goo, Young-Min;Kim, Tae-Won;Cho, Kwang-Soo;Cho, Hyeon-Seol;Yun, Dae-Jin;Chung, Woo-Sik;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.62-68
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    • 2011
  • Calmodulin (CaM), a $Ca^{2+}$ binding protein in eukaryotes, mediates cellular $Ca^{2+}$ signals in response to a variety of biotic and abiotic external stimuli. The $Ca^{2+}$-bound CaM transduces signals by modulating the activities of numerous CaM-binding proteins. As a CaM binding protein, AtCBP63 ($\b{A}$rabidopsis thaliana $\b{C}$aM-binding protein $\underline{63}$ kD) has been known to be positively involved in plant defense signaling pathway. To investigate the pathogen resistance function of AtCBP63 in potato, we constructed transgenic potato (Solanum tuberosum L.) plants constitutively overexpressing AtCBP63 under the control of cauliflower mosaic virus (CaMV) 35S promoter. The overexpression of the AtCBP63 in potato plants resulted in the high level induction of pathogenesis-related (PR) genes such as PR-2, PR-3 and PR-5. In addition, the AtCBP63 transgenic potato showed significantly enhanced resistance against a pathogen causing bacterial soft rot, Erwinia carotovora ssp. Carotovora (ECC). These results suggest that a CaM binding protein from Arabidopsis, AtCBP63, plays a positive role in pathogen resistance in potato.

Regulation of Arabidopsis Circadian Clock by De-Etiolated 1 (DET1) Possibly via Histone 3 Acetylation (H3Ac) (히스톤 3 아세틸화(H3Ac)를 통한 De-Etiolated 1 (DET1)의 애기장대 생체시계 조절)

  • Song, Hae-Ryong
    • Journal of Life Science
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    • v.22 no.8
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    • pp.999-1008
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    • 2012
  • The circadian clock is a self-sustaining 24-hour timekeeper that allows organisms to anticipate daily-changing environmental time cues. Circadian clock genes are regulated by a transcriptional-translational feedback loop. In Arabidopsis, LATE ELONGATED HYPOCOTYL (LHY) and CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1) transcripts are highly expressed in the morning. Translated LHY and CCA1 proteins repress the expression of the TIMING OF CAB EXPRESSION 1 (TOC1) transcripts, which peaks in the evening. The TOC1 protein elevates the expression of the LHY and CCA1 transcripts, forming a negative feedback loop that is believed to constitute the oscillatory mechanism of the clock. In mammals, the transcription factor protein CLOCK, which is a central component of the circadian clock, was reported to have an intrinsic histone acetyltransferase (HAT) activity, suggesting that histone acetylation is important for core clock mechanisms. However, little is known about the components necessary for the histone acetylation of the Arabidopsis clock-related genes. Here, I report that DET1 (De-Etiolated1) functions as a negative regulator of a key component of the Arabidopsis circadian clock gene LHY in constant dark phases (DD) and is required for the down-regulation of LHY expression through the acetylation of histone 3 (H3Ac). However, the HATs directly responsible for the acetylation of H3 within LHY chromatin need to be identified, and a link connecting the HATs and DET1 protein is still absent.

Effects of Frequency of Meals on Energy Utilization and Body Composition of Sheep Ingesting Diets of Equal Amount (급식회수(給食回數)가 면양(緬羊)의 열량대사(熱量代謝) 및 조직(組織)의 화학적성분(化學的成分)에 미치는 영향)

  • Han, In-K.
    • Applied Biological Chemistry
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    • v.8
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    • pp.21-28
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    • 1967
  • Two experiments with 32 sheep were conducted to study the effects of feeding the same amount of diet per day at different meal frequencies on ration digestibility, energy utilization, rate of gain, body composition and efficiency of gain. The results obtained are as follows: (1) The ingestion by sheep of the same amount of feed per day in 8 meals, 1 meal plus 7 ruminal inflations-deflations, and in 1 meal caused no different effect in the digestibility of the nutrients and energy, or the ME value of the diet. (2) Heat production per unit of metabolic size per unit of dietary intake was markedly lower for sheep ingesting 8 meals or administered 1 meal plus 7 ruminal inflations-deflations per day than for sheep fed 1 meal per day. (3) Body weight gain was significantly greater by sheep fed 8 meals per day or 1 meal plus 7 ruminal inflations-deflations than by those fed 1 meal per day. However, the gain in DM and energy of wool was not affected by frequency o( meals. (4) Sheep ingesting 8 meals or administered 1 meal plus 7 ruminal inflations-deflations per day gained body protein, fat and energy at a more rapid and efficient rate than sheep fed 1 meal per day. (5) Sheep fed 8 meals per day gained greater proportion of fat, protein and ash in the gained portion of the bodies than did 1 meal fed sheep. (6) An attempt was made to establish the possible explanations by which the frequency of ingesting meals exerts its effects.

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Isolation and Characterization of the nsdC Gene in Sexual Development of Aspergillus nidulans (Aspergillus nidulans의 유성분화에 관여하는 nsdC 유전자의 분리 및 분석)

  • Kim, Hye-Ryun;Han, Dong-Min
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.246-251
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    • 2006
  • A lot of mutants which cannot initiate sexual development were screened and several loci including nsdA, nsdB, nsdC, and nsdD were identified in homothallic ascomycetes Aspergillus nidulans. The NSD206, which has nsdC6 allele, showed typical phenotype of NSD (Never in sexual development) mutants. The nsdC gene was cloned by transforming NSDP697 ($nsdC^-$, $pryG^-$) with AMA1-NotI genomic library. The transforming library DNA recovered from several transformants showing wild phenotype carried about 10 kb genomic DNA insert. The DNA sequence of nsdC was analysed using GPS (Genome priming system). The nsdC gene has an open reading frame (ORF) of 1,929 bp encoding a putative polypeptide of 643 amino acids. The NsdC carries $C_2H_2C_2H_2C_2HC$ type zinc finger DNA binding domains in the middle of the polypeptide. A coiled-coil domain at its C terminus were also found. In nsdC6 allele, a single T insertion was occurred between 407-408 bp leading to the frameshift mutation and early termination of translation producing the truncated protein which has only 139 amino acids.

Implications of Streptomyces coelicolor RraAS1 as an activator of ribonuclease activity of Escherichia coli RNase E (Streptomyces coelicolor RraAS1의 Eschechia coli RNase E의 RNA 분해작용에 대한 활성제로서 기능 암시)

  • Heo, Jihune;Seo, Sojin;Lee, Boeun;Yeom, Ji-Hyun;Lee, Kangseok
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.243-248
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    • 2016
  • RNase E (Rne) is an essential enzyme involved in the processing and degradation of a large portion of RNAs in Escherichia coli. The enzymatic activity of RNase E is controlled by regulators of ribonuclease activity, namely, RraA and RraB. Gram-positive bacterium Streptomyces coelicolor also contains homologs of Rne and RraA, designated as RNase ES (Rns), RraAS1, and RraAS2. In the present study, we investigated the effect of S. coelicolor RraAS1 on the ribonucleolytic activity of RNase E in E. coli. Coexpression of RraAS1 with Rne resulted in the decreased levels of rpsO, ftsZ, and rnhB mRNAs, which are RNase E substrates, and augmented the toxic effect of Rne overexpression on cell growth. These in vivo effects appeared to be induced by the binding of RraAS1 to Rne, as indicated by the results of co-immunoprecipitation analysis. These results suggested that RraAS1 induces ribonucleolytic activity of RNase E in E. coli.

Characterization of ent-Kaurenoic Acid 13-Hydroxylase in Steviol Biosynthesis of Stevia rebaudiana Bertoni (Stevia rebaudiana Bertoni의 Steviol 생합성 효소 ent-Kaurenoic Acid 13-Hydroxylase의 특성)

  • Shibata, Hitoshi;Kim, Keun-Ki
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.501-507
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    • 1997
  • Chloroplasts isolated from Stevia rebaudiana Bertoni leaves contained an enzyme activity which catalyzed hydroxylation of ent-kaurenoic acid (ent-kaur-16-en-19-oic acid; ent-KA) to steviol (ent-13-hydroxy kaur-16-en-19-oic acid), the diterpenoid carboxylic alcohol which is the aglycone of sweet stevioside-related glycosides. $[^(14)C]-methylated$ ent-KA was used to localize ent-KA hydroxylase. $[^(14)C]-methyl-KA$ was most actively was transformed into methyl-steviol in chloroplast. The enzymatic activity was found in stroma fraction but not in thylakoid membrane in Stevia rebaudiana Bertoni. However, ent-KA 13-hydroxylase activity was not detected in stroma fraction of either Spinacia oleracea and Solidago altissima. The reaction products using $[^(14)C]-methyl-KA$ were purified and identified on TLC autoradiogram. The hydroxylation of ent-KA from stromal protein to form steviol required NADPH and oxygen. FAD and riboflavin stimulated the enzyme activity 1.5-and 1.7-fold, respectively. It also turned out that the activity of this enzyme using methyl-KA as a substrate was 16.7% that of ent-KA. The purified ent-KA 13-hydroxylase did not act on t-cinnamic acid, 4-hydroxyphenyl acetic acid, choline and resorcinol, known as monooxygenase and hydroxylase substrates.

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Molecular Cloning and Nucleotide Sequencing of a DNA Clone Encoding Arginine Decarboxylase in Rice (Oryza sativa L.) (벼의 arginine decarboxylase DNA clone의 재조합 및 염기서열 분석)

  • Hong, Sung-Hoi;Jeung, Ji-Ung;Ok, Sung-Han;Shin, Jeong-Sheop
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.112-117
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    • 1996
  • Arginine decarboxylase (ADC) is the first enzyme in one of the two pathways of diamine putrescine biosynthesis in plants. The genes encoding ADC have previously been cloned from Escherichia coli, oat and tomato genome. Two degenerate oligonucleotides (17-mer) corresponding to two conserved regions of ADC were used as primers in polymerase chain reaction of rice (Oryza sativa L.) genomic DNA, and an approximately 1.0 kbp fragment was obtained. This amplified PCR product showed an open reading frame which contains 1,022 bp of nucleotide sequences. This PCR product was cloned into pGEM-originated T vector and the short 500 bp PstI digested fragment was subcloned into pGEM-3zf(+/-) vectors to facilitate sequencing. The nucleotide sequence of this PCR product showed about 74% and 70% identity with the same regions of the oat and tomato ADC cDNA sequences, respectively. The predicted amino acid sequence exhibited 45% and 62% identity with oat and tomato ADC polypeptide fragments, respectively. The sequence similarities of 34%, 47% and 38% were previously reported in oat and E. coli, tomato and oat, and tomato and E. coli ADC amino acids, respectively. Therefore, similarities and identities between rice and oat or tomato are remarkably higher than those others of the previous reports. In the highly conserved regions in both the amino acid sequence and spacing regions among the sequences of these three, rice ADC open reading frame also has the exactly same regions with the striking similarity. RNA blot analysis showed that hnc is expressed as a transcript of approximately 2.5 kbP in the rice seedling leaf tissues.

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