• Title/Summary/Keyword: Agricultural technology center

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Expression and Purification of Ubiquitin-Specific Protease (UBP1) of Saccharomyces cerevisiae in Recombinant Escherichia Coli

  • Na, Kang-In;Kim, Myoung-Dong;Min, Won-Ki;Kim, Jeong-Ah;Lee, Woo-Jong;Kim, Dae-Ok;Park, Kyung-Moon;Seo, Jin-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.599-602
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    • 2005
  • Truncated form of UBP1, an ubiquitin-specific protease of Saccharomyces cerevisiae, was overexpressed in Escherichia coli. The hexahistidine residue $(His_6)$ was fused to the N-terminus of truncated UBP1 and the corresponding recombinant protein was purified with high yield by immobilized metal affinity chromatography. The truncated form of UBP1 protein was functional to cleave ubiquitinated human growth hormone as substrate. Effects of pH and temperature were investigated in order to optimize deubiquitinating reactions for the truncated UBP1. Optimum temperature and pH for the cleavage reaction were $40^{\circ}C$ and pH 8.0, respectively.

Quantitation of CP4 5-Enolpyruvylshikimate-3-Phosphate Synthase in Soybean by Two-Dimensional Gel Electrophoresis

  • KIM YEON-HEE;CHOI SEUNG JUN;LEE HYUN-AH;MOON TAE WHA
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.25-31
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    • 2006
  • Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

The three proline residues (P25, P242, and P434) of Agrobacterium CP4 5-enolpyruvylshikimate-3-phosphate synthase are crucial for the enzyme activity

  • Kang, Kyung-Su;Jin, Yong-Mei;Jeon, Hye-Sung;Park, Sang-Ryoung;Song, Dae-Geun;Lee, Joo-Young;Pan, Cheol-Ho;Kim, Min-Kyun
    • Plant Biotechnology Reports
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    • v.4 no.4
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    • pp.329-334
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    • 2010
  • Multiple sequence alignments showed that the prolines at the 25th, 129th, 153rd, 242nd, 322nd, and 434th amino acids in 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium sp. strain CP4 are strongly conserved in various prokaryotic EPSPS proteins. Single point mutations of the conserved prolines to alanine (P25A, P153A, P242A, P322A, and P434A) were introduced in the CP4 EPSPS in order to investigate the importance of the conserved prolines for the enzyme properties. The point mutations caused decreases in substrate binding affinity and catalytic efficiency as well as the glyphosate resistance, in general. Especially, the 25th and 242nd prolines located in the polypeptide hinges connecting top and bottom domains of CP4 EPSPS as well as the 434th proline at the C-terminus of the enzyme turned out to be crucial for the enzyme activity.

Genomic analysis of Sheldrake origin goose hemorrhagic polyomavirus, China

  • Wan, Chunhe;Chen, Cuiteng;Cheng, Longfei;Liu, Rongchang;Fu, Guanghua;Shi, Shaohua;Chen, Hongmei;Fu, Qiuling;Huang, Yu
    • Journal of Veterinary Science
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    • v.19 no.6
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    • pp.782-787
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    • 2018
  • Goose hemorrhagic polyomavirus (GHPV) is not a naturally occurring infection in geese in China; however, GHPV infection has been identified in Pekin ducks, a domestic duck species. Herein, we investigated the prevalence of GHPV in five domestic duck species (Liancheng white ducks, Putian black ducks, Shan Sheldrake, Shaoxing duck, and Jinyun Sheldrake) in China. We determined that the Jinyun Sheldrake duck species could be infected by GHPV with no clinical signs, whereas no infection was identified in the other four duck species. We sequenced the complete genome of the Jinyun Sheldrake origin GHPV. Genomic data comparison suggested that GHPVs share a conserved genomic structure, regardless of the host (duck or geese) or region (Asia or Europe). Jinyun Sheldrake origin GHPV genomic characterization and epidemiological studies will increase our understanding of potential heterologous reservoirs of GHPV.

Identification and Functional Analysis of Vibrio vulnificus SmcR, a Novel Global Regulator

  • Lee, Jeojng-Hyun;Rhee, Jee-Eun;Park, U-Ryung;Ju, Hyun-Mok;Lee, Byung-Cheol;Kim, Tae-Sung;Jeong, Hye-Sook;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.325-334
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    • 2007
  • Recently, quorum sensing has been implicated as an important global regulator controlling the production of numerous virulence factors such as capsular polysaccharides in bacterial pathogens. The nucleotide and deduced amino acid sequences of smcR, a homolog of V. harveyi luxR identified from V. vulnificus ATCC29307, were analyzed. The amino acid sequence of SmcR from V. vulnificus was 72 to 92% similar to those of LuxR homologs from Vibrio spp. Functions of SmcR were assessed by the construction of an isogenic mutant, whose smcR gene was inactivated by allelic exchanges, and by evaluating its phenotype changes in vitro and in mice. The disruption of smcR resulted in a significant alteration in biofilm formation, in type of colony morphology, and in motility. When compared with the wild-type, the smcR mutant exhibited reduced survival under adverse conditions, such as acidic pH and hyperosmotic stress. The smcR mutant exhibited decreased cytotoxic activity toward INT 407 cells in vitro. Furthermore, the intraperitoneal $LD_{50}$ of the smcR mutant was approximately $10^2$ times higher than that of parental wild-type. Therefore, it appears that SmcR is a novel global regulator, controlling numerous genes contributing to the pathogenesis as well as survival of V. vulnificus.

A Study on the Job-stress of Agricultural Extension Agents (농촌지도요원(農村指導要員)의 직무(職務)스트레스에 관한 연구(硏究))

  • Lee, Hee-Yul;Cho, Young-Man
    • Journal of Agricultural Extension & Community Development
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    • v.6 no.1
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    • pp.25-34
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    • 1999
  • This study was aimed 1) to determine the job-stress level of agricultural extension ants, 2).to analyze job-stress and its related variables. Data were collected from 512 extension ants who work at agricultural technology center in Pusan and Kyungnam province, 462 subjects were analyzed. SAS program was used. The major findings of this study were as follows: 1) the score of job-stress was low and that of job satisfaction was high,; 2) the degree of role overload of 40 year-old agents who have worked for over 20 years was high,; 3) the degree of role ambiguity of 20 year-old who have worked below 15 years was high,; 4) the degree of job-stress perception of 40 year-old agents was high,; 5) the degree of job satisfaction of among agents who finished graduate course and who are charge of section was high.

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Investigation of morphological changes of HPS membrane caused by cecropin B through scanning electron microscopy and atomic force microscopy

  • Hu, Han;Jiang, Changsheng;Zhang, Binzhou;Guo, Nan;Li, Zhonghua;Guo, Xiaozhen;Wang, Yang;Liu, Binlei;He, Qigai
    • Journal of Veterinary Science
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    • v.22 no.5
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    • pp.59.1-59.13
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    • 2021
  • Background: Antimicrobial peptides (AMPs) have been identified as promising compounds for consideration as novel antimicrobial agents. Objectives: This study analyzed the efficacy of cecropin B against Haemophilus parasuis isolates through scanning electron microscopy (SEM) and atomic force microscopy (AFM) experiments. Results: Cecropin B exhibited broad inhibition activity against 15 standard Haemophilus parasuis (HPS) strains and 5 of the clinical isolates had minimum inhibition concentrations (MICs) ranging from 2 to 16 ㎍/mL. Microelectrophoresis and hexadecane adsorption assays indicated that the more hydrophobic and the higher the isoelectric point (IEP) of the strain, the more sensitive it was to cecropin B. Through SEM, multiple blisters of various shapes and dents on the cell surface were observed. Protrusions and leakage were detected by AFM. Conclusions: Based on the results, cecropin B could inhibit HPS via a pore-forming mechanism by interacting with the cytoplasmic membrane of bacteria. Moreover, as cecropin B concentration increased, the bacteria membrane was more seriously damaged. Thus, cecropin B could be developed as an effective anti-HPS agent for use in clinical applications.

Isolation of Xanthine Oxidase Inhibitors from Ginkgo biloba Leaves-Derived Components

  • Baek, Bong-Rae;Kim, Moo-Key;Lee, Sung-Eun;Hwang, Young-Hee;Lee, Hoi-Seon
    • Preventive Nutrition and Food Science
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    • v.7 no.1
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    • pp.18-21
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    • 2002
  • The extract of Ginkgo biloba leaves was measured for inhibitory activity against xanthine oxidase. Aceton extract of G. biloba leaves showed strong inhibitory activity. Inhibitory activities of the fractionated extract were in the order of water > ethyl acetate fractions. Two fractions exhibiting strong inhibitory activities ware further purified via repeated silica gel, Amberlite IRN-78, Polyclar AT, and Sephadex LH-20 column chromatographies. Active components were isolated and identified through $^1$H-NMR and $^{13}$ C-NMR. The compounds were characterized as kaempferol 3-Ο-$\alpha$-(6$^{"′}$-p-coumaroylglucosyl-$\beta$-1,4-rhamnoside), and quercertin 3-Ο-$\alpha$-(6$^{′}$-p-coumaroylglucosyl-$\beta$-1,4-rhamnoside).

Lycopene Content and Fruit Morphology of Watermelon (Citrullus lanatus) Germplasm Collections

  • Jae-Jong Noh;On-Sook Hur;Na-Young Ro;Jae-Eun Lee;Ae-Jin Hwang;Bit-Sam Kim;Ju-hee Rhee;Jung Yoon Yi;Ji Hyun Kim;Ho-Sun Lee;Jung-Sook Sung;Myung-Kon Kim;Awraris Derbie Assefa
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.08a
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    • pp.32-32
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    • 2020
  • High-quality and high-phytonutrient watermelon fruits have strong market opportunities besides their health related benefits. Hence, investigating quality and nutritional related traits of watermelon genetic resources could provide important baseline data in breeding for increased lycopene content thereby increasing the marketability of watermelon. To this end, we have examined some fruit morphological traits and lycopene content of 105 genetic resources. The morphological characters were recorded on the field and inside laboratory and lycopene was measured using spectrophotometric and HPLC methods. Watermelon fruits have shown a diverse morphological characters. Red and pink fleshed fruits dominated in the entire collections. Fruits with higher thickness of rind were found to exhibit less soluble solid content (SSC). Korean origin fruits were characterized by intermediate SSC while USA, RUS, TJK, TKM, TWN, and URY originated fruits had the highest SSC. The lycopene content varied between 41.37 and 182.82 ㎍/g, 2.81 and 163.72 ㎍/g, and 3.54 and 255.47 ㎍/g using HPLC, UV-Vis, and microplate reader instrumnets, respectively. Red- and pink-fleshed fruits had the highest levels of lycopene content compared to the yellow- and orange-fleshed. Lycopene content had a significant positive correlation with SSC, however, no correlations were detected between lycopene and other quantitative fruit morphological characters.

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