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Use of the Yeast 1.5-Hybrid System to Detect DNA-Protein-Protein Interaction

  • Kim, Sook-Kyung;Han, Jin-Hee
    • Journal of Microbiology
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    • v.38 no.2
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    • pp.113-116
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    • 2000
  • Escherichia coli F plasmid partition apparatus is composed of two trans-acting proteins (SopA and SopB) and one cis-acting DNA sequence (sopC). The SopB-sopC complex has been suggested to serve a centromere-like function through its interaction with chromosomally encoded proteins which remain to be identified. In this paper, we are introducing a new yeast 1.5-hybrid system which assembles the two-hybrid and one-hybrid system as a mean to find and additional component of the F plasmid partition system, interacting with DNA (sopC)-bound SopB protein. The results indicates that this system is a promising one, capable of selecting an interacting component.

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Blood Component Change in Rat by Lipopolysaccharide and Cell Wall Protein-A from Vibrio vulnificus, E. coli, and S. typhimurium

  • Lee, Bong-Hun
    • Journal of Life Science
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    • v.10 no.2
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    • pp.9-11
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    • 2000
  • Lipopolysaccharide (LPS) and cell wall protein-A (CWP-A) were extracted from the cell wall of Vibrio vulnificus, Escherichia coli and Salmonella typhimurium. LPSs and CWP-As were injected into rat and the changes of the following blood components were examined. The change of the number of white blood cell (WBC), red blood cell (RCB), platelet (PLT), blood urea nitrogen (BUN) and blood glucose in rat blood and interferon (IFN) activity change by LPS and CWP-A were measured. WBC, RETI, PTT, and BUN were increased and RBC and blood glucose were increased slightly, but PLT was decreased.

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Changes In Protein and Its Components of High and Low Protein Varieties during Seed Development and Maturation of Soybeans (대두(大豆) 고저단백질(高低蛋白質) 품종(品種)의 성숙중(成熟中) 종실(種實)의 발육(發育)과 단백질(蛋白質) 및 그 구성분(構成分)의 변화(變化))

  • Lee, Jong Suk
    • Korean Journal of Agricultural Science
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    • v.5 no.2
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    • pp.56-67
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    • 1978
  • This study was conducted to characterize comparatively the accumulative patterns of protein and oil, temporal changes in electrophoretic components of proteins during seed development and maturation for the soybean varieties with high, medium and low protein contents. 1. The dry matter of the developing seed increases slowly for the first 22 days after flowering, followed by rapid linear increase for 20 to 30 days and further slow increase for 5 to 15 days attaining its maximum. During the period 12 to 27 days after flowering the protein content of seed increases rapidly while oil content increases rapidly. Following this period of rapid changes, there was period of slow increase until 40 to 47 days after flowering and no seizable further change in the content of both protein and oil. 2. The high protein variety, Saikai # 20, was characterized by shorter period and lower rate of decrease in protein content during the early period, followed by longer period and higher rate of increase in protein content, with earlier stop of oil accumlation during the seed development. 3. The low protein and high oil variety, Shelby, was characterized by longer period of decrease in protein content and shorter period of increase in protein content in contrast to the longer period of slow oil increase during seed development. 4. The temporal pattern of protein component accumulation during seed development was distinctly different among varieties differing in protein content. The time of distinct appearance of all the protein components identifiable in the matured seeds was in accordance with the end of d crease in the protein content of seed. A component having Rm of 0.03 which was absent in the matured seeds was identifiable during the first 17 days after flowering. 5. The high protein variety, Saikai # 20, had much higher compositioral ratio of the component a from the early days of seed development and it continued to increase until 47 days after flowering, while the increase in the composition of the component a stopped as early as 27 days after flowering in the other lower protein varieties. 6. The composition of the component b increased during the period from 17 to 42 days after flowering in all the varieties tested, but the rate of increase during the period was lowest in the high protein variety, Saikai # 20.

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Biochemical Changes in Muscle Protein of Squid Sikhae during Fermentation -Effects of Temperature and Moisture Content- (오징어 식해 숙성중 단백질 화학적 변화 -온도 및 수분함량의 영향-)

  • Lee, Nam-Hyouck;Oh, Se-Wook;Kim, Young-Myoung
    • Korean Journal of Food Science and Technology
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    • v.28 no.2
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    • pp.292-297
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    • 1996
  • In this study, biochemical changes of muscle protein in squid Sikhae prepared at various temperatures and moisture content were investigated. The pH values in squid Sikhae decreased rapidly with increase in titratable acidity during fermentation at high temperature and moisture content. Growth rate of lactic acid bacteria increased rapidly during fermentation at high temperature and moisture content. Changes in muscle protein subunit were analyzed by SDS-PAGE; myosin heavy chain component disappeared completely during preparation of squid Sikhae and actin component decreased slowly during fermentation. Therefore, it was suggested that myosin heavy chain component in squid muscle protein was autolyzed at the incipient for-mentation time and actin component was degraded by acidic protease in muscle and protease produced from microorganisms during fermentation.

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Physicochemical Properties of Recominant Hepatitis B Surface Antigen Expressed in Mammalian Cell(C127)

  • Lee, Young-Soo;Kim, Byong-Kak;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.5
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    • pp.521-526
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    • 1998
  • The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

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Lycium barbarum Polysaccharide Inhibits Lipid Oxidation and Protein Degradation in Tan Sheep Meatballs during Frozen Storage

  • Yu, Jiangyong;Guo, Mei;Liu, Guishan;Zhang, Jingjing;Fan, Naiyun;Li, Xiaorui;Sun, Yourui;Yuan, Jiangtao;Huang, Rui
    • Food Science of Animal Resources
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    • v.42 no.4
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    • pp.580-592
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    • 2022
  • The aim of the present study was to evaluate the effectiveness of Lycium barbarum polysaccharide (LBP) on lipid oxidation and protein degradation in Tan sheep meatballs during the frozen period. The meatballs were treated with LBP at 0.01%, 0.02%, and 0.03% and stored at -18±1℃ for 0, 3, 6, 9, and 12 weeks. The effects of LBP treatment were investigated using the contents of total volatile basic nitrogen (TVB-N), texture profile (TP), thiobarbituric acid reactive substances (TBARS), colour, and pH values, compared with 0.02% butylated hydroxytoluene treatment and the blank control. The results showed that LBP treatment significantly decreased TBARS content compared with the control, which confirmed LBP to be a highly effective component in preventing lipid oxidation of Tan sheep meatballs during frozen storage, and protein degradation in Tan sheep meatballs had a significant inhibition effect because of TVB-N value reduction. In addition, the colour, TP and pH values of meatballs treated with LBP were improved dramatically. To further determine the quality changes of the blank control and all treated groups during storage, the comprehensive score evaluation equation based on principal component analysis was obtained: Y=0.51632Y1+0.29589Y2 (cumulative contribution rate=81.221%), and the 0.02% LBP-treated group had a higher comprehensive score than the other groups, and the quality of LBP-treated meatballs was better as well. In summary, LBP may reduce or inhibit lipid oxidation and protein degradation, and enhance overall quality and shelf-life in prepared meat products.

Enhanced Production of Recombinant Protein in Escherichia coli Using Silkworm Hemolymph

  • Kim Ji Eun;Kim Eun Jeong;Rhee Won Jong;Park Tai Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.353-356
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    • 2005
  • The effect of silkworm hemolymph on the expression of recombinant protein in Escherichia coli was investigated. The addition of silkworm hemolymph to the culture medium in­creased the production of recombinant $\beta$-galactosidase in E. coli. The production was dependent on the concentration of the added silkworm hemolymph, which increased 2-, 5-, and 8-fold in media supplemented with 1, 3, and $5\%$ silkworm hemolymph, respectively. To identify the effective component, the silkworm hemolymph was fractionated by gel filtration column chromatography. A fraction, with a molecular weight of about 30 K was identified as the effective component.

A comparative study of the major component of the protein secretion machinery, secY, in gram positive bacillus subtilis and gram negative escherichia coli. (그람 양성균인 Bacillus subtilis와 그람 음성균인 escherichia coli에서 protein secretion에 중요 역할을 하는 secY에 대한 비교 연구)

  • 서주원
    • The Microorganisms and Industry
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    • v.17 no.1
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    • pp.10-18
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    • 1991
  • A phylogenetic comparison of homologous protein can often supplement genetic and biochemical analysis by revealing conserved structures that are critical for function(Waugh et al., 1989). I therefore isolated a secY homologue from B. subtilis, a gram positive bacterium evolutionary distant from E. coli. The comparison and interplay between these two bacterial systems should contribute greatly to our understanding of the functions and interactions within systems evolved for protein translocation in both prokaryotic and eukaryotic organisms.

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Identification of Protein Kinases by Anti-phosphoserine/Phosphothreonine/Phosphotyrosine Antibody Immunoaffinity Column Chromatographies in Streptomyces griseus. (Anti-Phosphoserine/Phosphothreonine/Phesphotyrosine Antibody Immunoaffinity Column Chromatography를 이용한 Streptomyces griseus의 인산화 단백질 동정)

  • Cheong, Yong-Hoon;Kim, Jong-Hee
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.112-117
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    • 2007
  • Protein kinases play very important role for maintaining viability in prokaryote and eukaryote. The metabolism of prokaryotic cell is generally regulated by bacterial two-component regulatory systems that are composed of histidine and asparitic acid kinases, however, some eukaryotic signal transduction system such as, serine and threonine kinases, have been also found to be involved in the regulation of morphogenesis and physiological differentiation in Streptomyces. Streptomyces griseus, a streptomycin producer, was expected to have varlous types of eukaryotic-type serine/threonine protein kinases, controlling morphogenesis. Thus, many steps of chromatographies were applied to isolate serine and threonine kinases from S. griseus IFO13350. The immunoaffinity steps using anti-phosphoserine, anti-phosphothreonine, and anti-phosphotyrosine agarose column chramatographies were successfully introduced to identify eukaryotic protein kinases from S. griseus IFO13350. Eight proteins with the expected molecular weight of 14, 29, 31, 35, 40, 52, 56, and 60 kDa, were identified on SDS-PAGE, and the their kination activity was confirmed by nonradioactive protein kination assay using FITC-labeled peptide as the substrate.

Correlation between Nutrient Components of Grass Roughages Mainly Used in Korea (국내 주요 화본과 조사료의 영양성분 간 상관관계)

  • Kim Myeong Hwa;Seo In Jun;Kim Joon Sun;Lee Sang Rak
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.3
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    • pp.169-176
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    • 2005
  • To evaluate the possibility of predicting a specific nutrient component, which nay be difficult in analysis, from a component of nutrient relatively easy to analyze, six grass roughage sources (bermuda grass hay, klein grass hay, oat hay, orchard grass straw, tall fescue straw and timothy hay) mainly used in Korea were analyzed for the heat-stable $\alpha-amylase$ treated neutral detergent fiber (aNDF), acid detergent fiber (ADF), acid detergent lignin (ADL), neutral detergent insoluble crude protein (NDICP) and acid detergent insoluble crude protein (ADICP) as well as proximate analytical components, and correlation coefficients between nutrient components were calculated. Some chemical components appeared to have significant correlation (p<0.05) through Pearson's correlation coefficients (i.e. between crude protein (CP) and NDICP in oat hay, orchard grass straw and tall fescue, between CP and ADICP in oat hay and timothy hay, between crude fiber (CF) and aNDF in klein grass hay, tall fescue straw and timothy hay, between CF and ADF in oat hay, tall fescue straw and timothy hay, and between CF and ADL in orchard pass straw and timothy hay). It is therefore suggested that it would be worthy to predict a specific nutrient component, which may be difficult in analysis such as aNDF, ADF, ADL, NDICP and ADICP, from a component of nutrient relatively easy to analyze such as CP and CF. However, it would be needed to improve the correlation coefficients, more samples should be measured.