• Title/Summary/Keyword: E. coli system

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Trichomonas vaginalis Adhesion Protein 33: A Useful Target for Diagnosis of T. vaginalis

  • Joo Kyung Bok;Rho Pyong-Ui;Park Seung Kyu;Kim Jae Won;Lee Kyu Jae;Ryang Yong Suk
    • Biomedical Science Letters
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    • v.11 no.1
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    • pp.15-22
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    • 2005
  • Trichomoniasis is a sexually transmitted disease induced by Trichomonas vaginalis, a parasitic protozoan. The symptoms of trichomoniasis are rarely appeared that the infections are distributed worldwide from underdeveloped to developed countries. The diagnosis of trichomoniasis is mainly taken by wet smear following microscopic examination, of which the diagnostic accuracies are poor and varies with the clinicians' experiences. Therefore, more exact and convenient diagnostic methods for T. vaginalis are required. Here, we cloned and expressed recombinant T. vaginalis adhesion protein 33 (rTvAP33) using an E. coli expression system. rTvAP33 was then immunized to rabbit and BALB/c mice for the production of anti-rTvAP33 antibodies. Sandwich ELISA using these antibodies detected T. vaginalis cultured in TYM broth supplemented with ferrous ions. Vagina-parasitizing microorganisms showed low cross-reactivities in this system. These results suggest that Tv AP33 is a good diagnostic target for the detection of TvAP33-expressing T. vaginalis.

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Detection of Iron Nanoparticles using Nuclear Magnetic Resonance Relaxometry and Inverse Laplace Transform

  • Kim, Seong Min
    • Journal of Biosystems Engineering
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    • v.39 no.4
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    • pp.345-351
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    • 2014
  • Purpose: Rapid detection of bacteria is very important in agricultural and food industries to prevent many foodborne illnesses. The objective of this study was to develop a portable nuclear magnetic resonance (NMR)-based system to detect foodborne pathogens (E. coli). This study was focused on developing a method to detect low concentrations of magnetic nanoparticles using NMR techniques. Methods: NMR relaxometry was performed to examine the NMR properties of iron nanoparticle mixtures with different concentrations by using a 1 T permanent magnet magnetic resonance imaging system. Exponential curve fitting (ECF) and inverse Laplace transform (ILT) methods were used to estimate the NMR relaxation time constants, $T_1$ and $T_2$, of guar gum solutions with different iron nanoparticle concentrations (0, $10^{-3}$, $10^{-4}$, $10^{-5}$, $10^{-6}$, and $10^{-7}M$). Results: The ECF and ILT methods did not show much difference in these values. Analysis of the NMR relaxation data showed that the ILT method is comparable to the classical ECF method and is more sensitive to the presence of iron nanoparticles. This study also showed that the spin-spin relaxation time constants acquired by a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence are more useful for determining the concentration of iron nanoparticle solutions comparwith the spin-lattice relaxation time constants acquired by an inversion recovery pulse sequence. Conclusions: We conclude that NMR relaxometry that utilizes CPMG pulse sequence and ILT analysis is more suitable for detecting foodborne pathogens bound to magnetic nanoparticles in agricultural and food products than using inversion recovery pulse sequence and ECF analysis.

Developing New Mammalian Gene Expression Systems Using the Infectious cDNA Molecular Clone of the Japanese Encephalitis Virus

  • Yun Sang-Im;Choi Yu-Jeong;Park Jun-Sun;Kim Seok-Yong;Lee Young-Min
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2003.05a
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    • pp.83-86
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    • 2003
  • Major advances in positive-sense RNA virus research have been facilitated by the development of reverse genetics systems. These systems consist of an infectious cDNA clone that encompasses the genome of the virus in question. This clone is then used as a template for the subsequent synthesis of infectious RNA for the generation of synthetic viruses. However, the construction of infectious cDNA for the Japanese encephalitis virus (JEV) has been repeatedly thwarted by the instability of its cDNA. As JEV is an important human pathogen that causes permanent neuropsychiatric sequelae and even fatal disease, a reliable reverse genetics system for this virus is highly desirable. The availability of this tool would greatly and the development of effective vaccines as well as facilitate studies into the basic biology of the virus, including the molecular mechanisms of viral replication, neurovirulence, and pathogenesis. We have successfully constructed a genetically stable infectious JEV cDNA containing full-length viral RNA genome. Synthetic RNA transcripts generated in vitro from the cDNA were highly infectious upon transfection into susceptible cells, and the cDNA remained stable after it had been propagated in E. coli for 180 generations. Using this infectious JEV cDNA, we have successfully expressed a variety of reporter genes from the full-length genomic and various subgenomic RNAs in vitro transcribed from functional JEV cDNAS. In summary, we have developed a reverse genetics system for JEV that will greatly facilitate the research on this virus in a variety of different fields. It will also be useful as a heterologous gene expression vector and aid the development of a vaccine against JEV.

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A Study about Microbiological Quality and Safety Control of a Central Commissary School Foodservice System in Daejeon City Area (대전지역 도시형 공동조리교 급식의 미생물적 품질관리에 관한 연구)

  • Kim, Heh-Young;Jeong, Hyo-Jin
    • Journal of the Korean Society of Food Culture
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    • v.10 no.1
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    • pp.67-74
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    • 1995
  • This study researched microbial change of quality according to the various phases of product flow of cooked pea and rice, cold cucumber and seaweed soup, soybean sprouts japchae feeding urban type of a commissary school and a satellite school in Daejeon area, also it suggested the possibility that the central commissary foodservice system can be established and utilized more developmental to identify its food of variation of temperature and state of safety unitl 3 hours after cooking for the case of delay of distribution and holding because of the satellite school of geographical location and traffic problem. The critical Control Points identified for each category of menu items were: Boiled pea and rice: inadequate distribution, holding and storing before assembly; Cold cucumber and seaweed soup: pre-preparation and post-preparation after cooking; Soybean sprouts japchae: Pre-preparation, post-preparation and storing. As the result of observation of the variation of temperature and microbial safety according to the delay of distribution and holding for each food, all of them were relatively safe until 3 hours after cooking, but cold cucumber and seaweed soup being stored for 3 hours, the value of E. coli is $10^3$ CFU/g. The variation of temperature was more extreme in soybean sprouts japchae than cooked pea and rice and cold cucumber and seaweed soup. It was proved that the stainless container was excellent and that adequate holding container should be used.

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Effect of Adjuvants on Antibody Titer of Synthetic Recombinant Light Chain of Botulinum Neurotoxin Type B and its Diagnostic Potential for Botulism

  • Jain, Swati;Ponmariappan, S.;Kumar, Om;Singh, Lokendra
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.719-727
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    • 2011
  • Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

Measurement of Delivery Service Environment for Cold Chain EPS Packaging System of Fresh Food (신선식품 콜드체인 EPS 패키징 시스템의 택배 유통환경 계측)

  • KORAKOT, CHAROENSRI;Kim, SY;Shin, YJ;Jung, HM;Park, JM
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.28 no.1
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    • pp.67-73
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    • 2022
  • The food cold chain refers to a technology and distribution supply chain applied to maintain a constant temperature suitable for the product from production (harvest) to delivery to consumers. In particular, in Korea, the insulation material used in the food cold chain is mostly EPS (Expanded Polystyrene), which is used as a transport container for various food cold chains. However, according to the government's eco-friendly policy, companies charge environmental contributions to the use of EPS, but due to its low price and convenience of handling, it is still used as a container for delivering food. In this study, in order to measure the domestic delivery environment of general refrigerated foods, changes in impact, temperature, and humidity during transport of the EPS packaging system containing foods and ice pack refrigerants were measured. As a result, there were 2?3 sections in which a high impact force of 40 G or more was generated during transport. This can cause damage to the product and EPS container. The difference in temperature and humidity changes by parcel transport routes is more than 30%, so it is necessary to present accurate standards for the domestic cold chain distribution environment. As a result of microbial experiments. the transportation period had a dominant effect on the increase in total viable count and E. coli count.

Characterization and Antimicrobial Activity of Lactic Acid Bacteria Isolated from Vaginas of Women of Childbearing Age (가임기 여성의 질에서 분리한 젖산 세균인 Lactobacillus plantarum UK-3의 특성 및 항균활성)

  • Ahn, Hye-Ran;So, Jae-Seong;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.308-315
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    • 2011
  • The purpose of this work was to examine the antimicrobial activity derived from the lactic acid bacterium, UK-3 isolated from the vaginas of women of childbearing age. Various physiological and biochemical properties of this strain were characterized. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was designated as Lactobacillus plantarum UK-3, and registered in GenBank as [JK266589]. Growth rate, production of organic acids (e.g., lactic acid and acetic acid), and pH during growth were monitored. The maximum concentrations of lactic acid and acetic acid were approximately 684.11 mM and 174.26 mM, respectively, and pH changed from 7.0 to 3.7 after 72 h of incubation. High performance liquid chromatography was used to confirm lactic acid and acetic acid production. Significant antimicrobial activity of the concentrated supernatant was demonstrated against various Gram-positive (e.g., Staphylococcus aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Neisseria species., Listeria monocytogenes), Gram-negative bacteria (e.g., Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis), and yeast (e.g., Candida albicans) by the plate diffusion method. As a result, the concentrated L. plantarum UK-3 cultures had lower acidity and inhibited the growth of all microorganisms tested, whereas the growth of L. acidophilus was not affected.

Investigation on Inhibitory Effect of ErmSF N-Terminal End Region Peptide on ErmSF Methyltansferase Activity In Vivo Through Development of Co-Expression System of Two Different Proteins in One Cell (서로 다른 두 단백질의 세포 내 동시 발현 체계의 개발을 통한 ErmSF에서 특이적으로 발견되는 N-Terminal End Region (NTER)을 포함하는 펩타이드의 생체내에서의 ErmSF 활성 억제 효과 검색)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.47 no.3
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    • pp.200-208
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    • 2011
  • Most problematic antibiotic resistance mechanism for MLS (macrolide-lincosamide-streptogramn B) antibiotics encountered in clinical practice is mono- or dimethylation of specific adenine residue at 2058 (E. coli coordinate) of 23S rRNA which is performed by Erm (erythromycin ribosome resistance) protein through which bacterial ribosomes reduce the affinity to the antibiotics and become resistant to them. ErmSF is one of the four gene products produced by Streptomyces fradiae to be resistant to its own antibiotic, tylosin. Unlike other Erm proteins, ErmSF harbors idiosyncratic long N-terminal end region (NTER) 25% of which is comprised of arginine well known to interact with RNA. Furthermore, NTER was found to be important because when it was truncated, most of the enzyme activity was lost. Based on these facts, capability of NTER peptide to inhibit the enzymatic activity of ErmSF was sought. For this, expression system for two different proteins to be expressed in one cell was developed. In this system, two plasmids, pET23b and pACYC184 have unique replication origins to be compatible with each other in a cell. And expression system harboring promoter, ribosome binding site and transcription termination signal is identical but disparate amount of protein could be expressed according to the copy number of each vector, 15 for pACYC and 40 for pET23b. Expression of NTER peptide in pET23b together with ErmSF in pACYC 184 in E. coli successfully gave more amounts of NTER than ErmSF but no inhibitory effects were observed suggesting that there should be dynamicity in interaction between ErmSF and rRNA rather than simple and fixed binding to each other in methylation of 23S rRNA by ErmSF.

A study on the Development of a Drying and Fermentation Process of Domestic Animal Manure;II. Demonstration of a Pig Manure Treatment System on a Farm (가축분(家畜糞) 건조(乾燥) , 발효(醱酵) 복합시설(複合施設) 개발(開發) 연구(硏究);II. 돈분(豚糞) 건조(乾燥), 발효(醱酵), 복합시설(複合施設) 실증시험(實證試驗))

  • Yun, Sun-Gang;Jung, Kwang-Yong;Park, Woo-Kun;Kwon, Sun-Ik;Park, Hong-Jae;Yoo, Sun-Ho
    • Korean Journal of Environmental Agriculture
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    • v.13 no.2
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    • pp.223-230
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    • 1994
  • A practical study on a drying and fermentation system equipped with a stirring machine operated mechanically, of pig manure was conducted to prove the efficiency of and practicability to an ordinary pig farm. The type of the drying bed was a round-shaped (r=3m) concrete structure and the stirring machine was adopted to stir and transfer dried pig manure to the fermentation tank. The dried pig manure was put into a fermentation tank ($V=18m^3$), which was aerated from pipe lines installed at the bottom. While water content of pig manure passing through a drying bed was remarkably reduced than before drying, the drying efficiency of this system decreased in winter. However, the temperature of pig manure piled up in the fermentation room in winter reached over $60^{\circ}C$ and excess water of pig manure was removed during the fermentation process. The reduction rate of water content of pig manure, to which dried pig manure was added as bulking material on the drying bed, was 52.1%, but when dried without bulking material it was only 19.7%. Although the content of $P_2O_5$ of dried pig manure was slightly higher than that of fresh pig manure, progressive changes in chemical composition between fresh and dried pig manure made no great difference. Among the contents of minerals of fresh and dried pig manure, CaO was the highest and the rest were in the decreasing order of $K_2O$, MgO, and $Na_2O$. Population density of E. coli and Streptococci of dried pig manure was reduced by 142 and 236 times that of fresh pig manure, respectively. The installation cost of this drying and fermentation system was 4,185,630 won (approximately 5,232 US $) and operating cost per year was 190,000 won (237.5US $) on the basis of self-labor condition.

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Desmutagenic Effect of Water Extract from Artemisia capillaris THUNB on the Mutagenicity of Benzo[a]pyrene (Benzo[a]pyrene의 변이원성에대한 인진쑥 물 추출물의 항돌연변이 효과)

  • 안병용
    • KSBB Journal
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    • v.15 no.4
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    • pp.331-336
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    • 2000
  • The antimutagenic activity of the extract of Artemisia capillaris THUNB on the mutagenicity induced by benzo(a)pyrene [B(a)P] in the presense of S9 mixture was studied using bacterial mutagenic assay system. Samples harvested in summer and autumn were extracted using ethanol and hot water. Among these extracts the water extract of summer sample had the strongest inhibitory effect against the mutagenenicity of B(a)P, The water extract of Artemisia capillaris THUNB was separated again into ethanol soluble and insoluble parts. The ethanol insoluble part(El) of water extract exhibited higher inhibition effects than the ethanol soluble part against the mutagenic activity of B(a)P. El showed dose-dependent activity on the mutagenicity of B(a)P in SOS Chromotest and Ames test. The 50% inbibition concentraction $(IC_{50}$ of El were $200{\mu}g/assay$ $600{\mu}g/plate$ and $800{\mu}b/plate$ in E. coil PQ37 S. typhimurium TA100 and TA98 respectively. El were showed desmutagenic effect but had no effect on the DNA repair system for B(a)P-induced mutagenesis. HPLC analysis showed that the formation of aflatoxin M1 by cytochrome P-450 1A1 known as playing an impotant role on B(a) P-induced mutagenicity was highly inhibited by El. Therefore we encluded that B(a)P-induced mutagencity can be reduced possible due to the interference of el with cytochrome P-450 1A1-dependent bioactivation.

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