• Title/Summary/Keyword: C coli

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Molecular Cloning and Expression of a Cu/Zn-Containing Superoxide Dismutase from Thellungiella halophila

  • Xu, Xiaojing;Zhou, Yijun;Wei, Shanjun;Ren, Dongtao;Yang, Min;Bu, Huahu;Kang, Mingming;Wang, Junli;Feng, Jinchao
    • Molecules and Cells
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    • v.27 no.4
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    • pp.423-428
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    • 2009
  • Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

Effect of Reusing Sawdust Fermented with Swine Excretion on the Growth Performance of Pigs (톱밥발효돈분 재이용이 돼지 성장에 미치는 영향)

  • 유용희;김형호;정일병;장병귀;곽정훈;박치호;한정대
    • Journal of Animal Environmental Science
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    • v.2 no.1
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    • pp.53-62
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    • 1996
  • This experiment was conducted to investigate the effect of the reuse of sawdust fermented with swine excretion as bed material on the growth performance of pigs. The sawdust which was already fermented with swine excretion in the pig house for eight months was transported to a fermentation facility for secondary fermentation. A total of 96 pigs with average 30kg of initial body weight were randomly assigned in the $2{\times}2$ factorial design with two levels(0%, 1.5%) of probiotics added for secondary fermentation and two levels(0%, 1.5%) of probiotics in feed. The results obtained are as follows : 1. Total-nitrogen(T-N), $K_2O$, total-carbon(T-C), and carbon/nitrogen(C/N) in sawdust bed showed no significant difference within treatments, but phosphate increased by 57% in average compared to the initial. 2. There was no significant difference in temperature in the sawdust bed treatments. 3. The internal parasite eggs detected were Trichuris suis, Strong. ransomi, Ascaris suum, Coccidia and Balantidium coli. 4. The utilization period of sawdust fermented with swine excretion was 52, 26, 16, 4, 5, 3 days, respectively, with increase of body weight. 5. Average daily gain and feed conversion were significantly improved by adding probiotics in the feed(P<0.05), but there was no difference between fermented sawdust with or without probiotics. 6. There was no significant difference in carcass weight and backfat thickness of pig among treatments(P>0.05).

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The Expression Patterns of Human Parkin in E. codi and Mammalian Cells (대장균과 포유류 세포 내에서 parkin의 발현 양상에 관한 연구)

  • Nam Min-Kyung;Park Hye-Min;Choi Ju-Youn;Park Hyo-Jin;Chung Kwang Chul;Kang Seong man;Rhim Hyangshuk
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.916-922
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    • 2005
  • Parkin, known as an E3 ubiquitin ligase, has essential role in protein quality control, and its severe dysfunction leads to neurodegenerative disorders. Human Parkin was excessively degraded when expressed in Escherichia coli under the conventional induction condition ($37^{\circ}C$ culture condition with 0.5 mM IPTG). To optimize the induction and culture conditions for recombinant human Parkin and develop a rapid method for the Parkin purification, we expressed Parkin by using PCEX system at the different culture temperatures and IPTC concentrations. The intact Parkin protein was purified to approximately $90\%$ purity with suitable amounts of protein under the optimal culture condition ($25^{\circ}C$E with 0.01 mM IPTG). Additionally, we constructed various parkin plasmids with different tagging systems and investigated their expression patterns in HEK293 cells. We found that the proteolytically sensitive site is localized within a ubiquitin-like domain of Parkin. This study developes a method for generating useful reagents to investigate biochemical properties of Parkin.

Cloning and Expression of Antifungal Protein (PR5) Genes from Hot Pepper (Capsicum annuum L.) (고추(Capsicum annuum)의 항균성 단백질(PR-5) 유전자의 클로닝과 발현 분석)

  • Park, Hae-Jin;Lee, Jung-Hoon;Yoon, Yong-Hwi;Kim, Hak-Yoon;Shin, Dong-Hyun;Lee, In-Jung;Kim, Dal-Ung;Kim, Kil-Ung
    • Journal of Life Science
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    • v.12 no.3
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    • pp.264-273
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    • 2002
  • We have isolated and artificially expressed three cDNA clones of Capsicum annuum PR5 genes for elucidating the antifungal activity against Phytophthora capsici which contracted a hot pepper root rot in field condition. Three divergent PR5 proteins from hot pepper were designated as CAPR5-1 and CAPR5-2 from susceptible cultivar (Subicho) as well as CAPR5-3 from resistant cultivar (CM331) in response to P. capsici. The cDNA similarity was found over 80% of identity among the three CAPR5s, and deduced amino acid sequence was characterized that all of CAPR5s contained 16 cysteine residues which possibly had a significant role in the structural formation. The result of genomic DNA blot showed that CAPR5-1 and CAPR5-2 existed as single copy in the Subicho genome. Three recombinant CPARs in E. coli were identified by SDS-PACE, and each expressed protein was treated on the PDA medium which contained cultured pathogens. Although three CAPR5 proteins did not affected the hyphal growth of Glomerella glycines and Colletotrichum fagenarium, CAPR5-1, CAPR5-2, and CAPR5-3 showed a specific antifungal activities against P. capsici.

Inhibitory Effect of Various Cereal and Bean Extracts on Carcinogenicity in vitro (곡류 및 두류 에탄올 추출물의 in vitro 발암 억제 효과 비교)

  • Choi, Yeong-Hee;Kang, Mi-Young;Nam, Seok-Hyun
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.964-969
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    • 1998
  • To investigated the anticarcinogenic activity of 70% ethanol extracts from various cereal in vitro, antimutagenic activity, inhibitory effect of DNA strand scission and tumor promotion were examined. The antimutagenic activity of the beans such as black bean and small red bean was generally higher than that of cereals examined. However inhibitory activity of 70% ethanolic extracts against DNA strand scission induced mitomycin C showed that millet, job's tear, black bean and soy bean among cereals and beans tested in this study inhibited effectively the DNA strand scission. Antioxidative activity of some cereal extracts determined by using linoleic acid model system showed that Job's tear, millet and black bean were higher antioxidative activity than other cereals and beans. Conventional short-term antipromoter assay system using activation of Epstein Barr virus (EBV) clearly demonstrated that sorghum, buckwheat, black bean and small red bean have inhibitory effects on promotion in cellular carcinogenesis.

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Biological and Chemical Hazards Factor Analysis for CCP(Critical Control Point) in Fried Process of Fried Noodles (유탕면류의 유탕공정 중 중요관리점(CCP)을 위한 미생물학적, 화학적 위해요소분석)

  • Kwon, Sang-Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.8
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    • pp.3578-3585
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    • 2012
  • The purpose of this study was to establish the critical limit at CCP (Critical Control Point) of HACCP (Hazard Analysis Critical Control Point) system for instant noodle and it was conducted at P company in Ichen(Gyeonggi-do), Korea. According to the CCP, Fried process were experimented to removal and decrease of microbiological and chemical hazards by measuring of each temperature and times. As a result, the standard plate count and pathogenic microorganism were not detected by fried processing (Temperature : $145{\pm}10^{\circ}C$, Time : $75{\pm}30$ sec). The acid value of chemical hazards produced by fried processing was able to manage, showed lower (0.2) than the legal limit (0.6). Air-borne bacterial examination results detected(3 CFU/mL, 3 CFU/mL) in the Frying Room and Steam Room. Therefore, the CCP-BC of fried process would be a great alternative to prevent and remove hazard analysis, such as general and pathogenic microorganism (E. coli O157:H7, B. cereus, Listeria monocytogenes, Salmonella spp, Sthaph. aureus etc), chemical hazard analysis. In conclusion, it suggested that HACCP plan was necessary for management standard and systematic approach in establishement of critical limit, solving the problem, method of verification, education and records management by fried processing.

Monoclonal antibody production for CP4 EPSPS detection assays (CP4 EPSPS 검출을 위한 단클론 항체 생산)

  • A-Mi Yoon;Il Ryong Kim;Wonkyun Choi
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.445-451
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    • 2021
  • In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

Preservation of Strawberries and Cucumbers Packaged by Low density polyethylene film impregnated with antimicmbial agent, Scutellariae baicalensis extract (황금추출물을 함유한 항균성 포장필름을 이용한 딸기와 오이의 저장효과)

  • 정순경;조성환
    • Food Science and Preservation
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    • v.9 no.3
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    • pp.271-276
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    • 2002
  • To develop a wrapping film, which suppresses the microbial decay through the storage and prolongs the selflife of fruits and vegetables, the antimicrobial packaging films were prepared and applied to the preservation of strtwberries and cucumbers. Low density polyethylene(LDPE) film of 50㎛ thickness was faricated with 1% of Scutellariae baicalensis extract. The LDPE film impregnated with Scutellariae baicalensis extract showed antimicrobial activity on the disk test against Bacillus cereus, Escherchia coli and Fusarium sp.. The antimicrobial film changed the color and light transmittance, but did not affect heat shrinkage, mechanical tensile strength and wattability. Strawberries and cucumbers were separately wrapped with packaging films in the state of closely-adhered packaging as well as modified atmosphere packaging(MAP). The wrapped strawberries and cucumbers were stored for 21 days at 5$\^{C}$ and for 40 days at l0$\^{C}$, respectively. For the packaged strawberries and cucumbers at 5$\^{C}$ and 10$\^{C}$, the LDPE film impregnated with Scutellariae baicalensis extract showed the reduced growth of total aerobic bacteria, molds and yeasts and did not give any negative effect on other quality attributes during storage in comparison with conttrol film without any additive.

Changes in Quality Characteristics of Seasoned Soy Sauce Treated with Superheated Steam and High Hydrostatic Pressure during Cold Storage (과열증기와 초고압 처리법을 적용한 간장 소스의 냉장저장 중 품질 특성 변화)

  • Choi, Yoon;Oh, Ji-Hye;Bae, In-Young;Cho, Eun-Kyoung;Kwon, Dae-Joong;Park, Hae-Won;Yoon, Sun
    • Korean journal of food and cookery science
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    • v.29 no.4
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    • pp.387-398
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    • 2013
  • Seasoned soy sauce is one of the popular seasoning sauces added to the Korean traditional foods such as Bulgogi, Galbi. However, industrially processed sauces have poor sensory quality because of heating treatment for sterilization. The purpose of this study was to develop seasoned soy sauce having fresh taste and good quality by applying superheated steam (SHS) and high hydrostatic pressure (HHP) technologies. To maintenance the sauce qualities, food materials such as apple, onion, and garlic were pretreated with SHS (heater $100^{\circ}C$, steam $280^{\circ}C$, 30 s~1 min 30 s) before mixing with other ingredients. During storage of 7 days, color, pH, and browning potential of SHS treated samples (apple, onion and garlic) did not change and also polyphenol oxidase was inactivated (p<0.05). The seasoned soy sauce including SHS treated materials was sterilized by thermal process ($85^{\circ}C$, 30min) or non-thermal process, HHP (550 MPa, $5{\sim}10^{\circ}C$, 3 min). In SHS+HHP treated sauce, salinity, sugar contents, lightness, viscosity did not change (p<0.05), and total viable cell counts were detected below 4 log cycle at $5^{\circ}C$ for 30 days. E.coli and B.cereus are not determined in all samples. In sensory evaluation, Bulgogi prepared with SHS+HHP treated sauce was more acceptable than others.

Studies for Antibiotic Free Chicken Production Using Water Extracts from Artemisia capillaris and Camellia sinensis (인진쑥 및 녹차 추출물을 이용한 무항생제 닭고기 생산 연구)

  • Kim, Dong-Wook;Kim, Ji-Hyuk;Kang, Geun-Ho;Kang, Hwan-Ku;Park, Sung-Bok;Park, Jae-Hong;Bang, Han-Tae;Kim, Min-Ji;Na, Jae-Cheon;Chae, Hyun-Suk;Choi, Hee-Chul;Suh, Ok-Suk;Kim, Sang-Ho;Kang, Chang-Won
    • Food Science of Animal Resources
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    • v.30 no.6
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    • pp.975-988
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    • 2010
  • Two experiments were conducted to determine whether water extracts from Artemisia capillaries (A. capillaries) and Camellia sinensis (C. sinensis) could be used as alternatives to antibiotic growth promoters in broiler feed. The experiment 1 was verified their chemical composition, extracts yields, total phenolic compounds concentration, antioxidant activity, antimicrobial activity, and chicken splenocytes proliferation through in vitro test. The extract yields of A. capillaries and C. sinensis were 26.5 and 16.8%, respectively. Total phenolic compounds concentrations of them expressed as gallic acid equivalent were 15.28 and 26.74 mg/mL, respectively. Electron donating abilities of them expressed as $SC_{50}$ showing 50% DPPH radical scavenging were 0.30 and 0.06 mg, respectively. Bacterial inhibitory rates of them against Escherichia coli, Staphylococcus aureus, and Salmonella Typhimurium were ranged from 42.1 to 52.3% and from 21.6 to 33.7%, respectively. And, these extracts increased proliferation of chicken splenocytes. Especially, A. capillaris was more excellent than Echinacea and Concanavalin A known as T-cell stimulator. The experiment 2 was investigated their effects on growth performance, relative organ weight, cecal microflora, blood biochemical parameters, and splenic cytokines mRNA expression in broiler chicks. Four hundred eighty 1-day-old male broiler chicks (Ross 308) were divided in to 4 treatment groups with 4 replicates of 30 birds in each group: NC (control, no antibiotics), PC (avilamycin, 10 ppm; salinomycin, 60 ppm), AC (A. capillaries, 100 ppm), and CS (C. sinensis, 100 ppm); treatments were administered through water supplementation. Final body weight was significantly higher in all treated groups than in NC (p<0.05). Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in NC (p<0.05). The relative weights and lengths of the small intestine were more significantly decreased in the PC and AC groups than in the other groups. Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in the NC group (p<0.05). The contents of total cholesterol, aspatate aminotransferase, and alanine aminotransferase in blood serum were more significantly decreased in all treated groups than in NC (p<0.05). In conclusion, these results suggested the possibility that these extracts could serve as alternatives for antibiotic growth promoters.