+

WO1990004053A1 - Dispositif d'electropolissage de surfaces - Google Patents

Dispositif d'electropolissage de surfaces Download PDF

Info

Publication number
WO1990004053A1
WO1990004053A1 PCT/DE1988/000626 DE8800626W WO9004053A1 WO 1990004053 A1 WO1990004053 A1 WO 1990004053A1 DE 8800626 W DE8800626 W DE 8800626W WO 9004053 A1 WO9004053 A1 WO 9004053A1
Authority
WO
WIPO (PCT)
Prior art keywords
bioluminescence
gene
microorganisms
housing
indicator
Prior art date
Application number
PCT/DE1988/000626
Other languages
German (de)
English (en)
Inventor
Robert Weber
Original Assignee
Siemens Aktiengesellschaft
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Siemens Aktiengesellschaft filed Critical Siemens Aktiengesellschaft
Priority to DE8888908666T priority Critical patent/DE3878697D1/de
Priority to PCT/DE1988/000626 priority patent/WO1990004053A1/fr
Priority to EP88908666A priority patent/EP0436528B1/fr
Publication of WO1990004053A1 publication Critical patent/WO1990004053A1/fr
Priority to US07/683,261 priority patent/US5135632A/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C25—ELECTROLYTIC OR ELECTROPHORETIC PROCESSES; APPARATUS THEREFOR
    • C25F—PROCESSES FOR THE ELECTROLYTIC REMOVAL OF MATERIALS FROM OBJECTS; APPARATUS THEREFOR
    • C25F7/00—Constructional parts, or assemblies thereof, of cells for electrolytic removal of material from objects; Servicing or operating
    • G—PHYSICS
    • G21—NUCLEAR PHYSICS; NUCLEAR ENGINEERING
    • G21F—PROTECTION AGAINST X-RADIATION, GAMMA RADIATION, CORPUSCULAR RADIATION OR PARTICLE BOMBARDMENT; TREATING RADIOACTIVELY CONTAMINATED MATERIAL; DECONTAMINATION ARRANGEMENTS THEREFOR
    • G21F9/00—Treating radioactively contaminated material; Decontamination arrangements therefor
    • G21F9/001—Decontamination of contaminated objects, apparatus, clothes, food; Preventing contamination thereof
    • G21F9/002—Decontamination of the surface of objects with chemical or electrochemical processes
    • G21F9/004—Decontamination of the surface of objects with chemical or electrochemical processes of metallic surfaces

Definitions

  • the invention relates to a method for the detection of toxic substances in a liquid or gaseous environment with the help of specifically sensitive and / or resistant microorganism strains which
  • All microorganisms are capable of emitting light (bio-luminescence) by introducing a plasmid-encoded luciferase gene.
  • bioluminescence of the indicator organisms enables changes in their metabolism or the loss of their viability as an effect of a toxic substance in the test medium to be detected quickly and with great sensitivity.
  • microorganisms for measuring critical loads caused by toxic substances (US Pat. No. 3,981,777) and for determining antibiotic concentrations (EP. No. 0200 226) has been established.
  • the applications are based on the detection of the growth of the test organisms by counting colonies, turbidity measurements, nephelometry, etc.
  • These measuring methods require the cultivation of large amounts of microorganisms, so that reliable measurements are only possible after can be carried out over a period of 16 to 72 hours. In contrast, the method described below allows the evaluation of measurement results after less than 2 hours.
  • Naturally occurring bioluminescent microorganisms are of marine origin and therefore require a high ionic strength in the test medium.
  • the production of the required osmolarity means a possibly falsifying intervention in the measurement.
  • the invention described here makes the methods ex- use experimental genetics to transform in principle any desired bacterial strain by introducing a specially constructed plasmid vector (pGL3, see below) into an organism capable of bioluminescence. While the principle of transforming a bacterial strain with a plasmid-encoded luciferase LUX gene has already been described (US Pat. No. 4,581,335), the present invention represents a decisive improvement: the expression of the LUX gene and the ability to bioluminescence is no longer permanent (constitutive), but can be switched on and off in a temperature-dependent manner.
  • the feature of adjustability is achieved by construction of a plasmid Vek ⁇ catalyst which both the luciferase gene complex of Vi • brio harveyi (lux genes A and B), and the C allele Ig57 of phage lambda repressor gene ( contains thermolabile gene product1).
  • the inducible bioluminescence achieved by this concept has two important advantages compared to permanently light-emitting test organisms:
  • the signal to noise ratio of the individual measurement is improved.
  • Another characteristic of the development according to the invention is the possibility of introducing additional resistance or hypersensitivity to specific toxic mediating genes in the plasmid vector outlined; this is made possible by molecular cloning of a corresponding gene into existing specific restriction endonuclease sites.
  • Measurement results take about ten times less time than conventional microbiological cultivation and measurement methods and
  • any suitable, recommended or prescribed in standardized reference tests bacterial strain for bioluminescence measurements can be set up.
  • E.coli ATCC 25922 may be mentioned as an example of a bacterial strain which belongs to the latter category; this strain is the official (WHO) reference strain for standardized antibiotic inhibition tests.
  • E. coli ATCC 25922 and other strains frequently used in clinical and industrial tests (E. coli K- and B; chi 1776 etc.) have been successfully converted by the applicant into the species capable of bioluminescence by introducing the plasmid pGL3 .
  • the regulation takes place by using the allele 857 of the lambda repressor in a temperature-dependent manner.
  • the repressor protein By increasing the temperature to above 37 ° C, the repressor protein is inactivated and the LUX gene complex is released for transcription in mRNA, which ultimately results in an increase in the bioluminescence activity of the indicator organism by more than three orders of magnitude.
  • the difference in the size of the bioluminescence signal between the repressed and transcribed state of the LUX gene complex - and thus the signal-to-noise ratio of the measurements - is more than a factor of ten in test organisms equipped with pGL3 better than those which are induced to bioluminescence by chemical induction of the lacZ gene (cf. US Pat. No. 4,581,335).
  • the plasmid pGL3 which was designed for the generation of controllable bioluminescence in microorganisms, was constructed by using conventional molecular biological cloning techniques; it contains:
  • Luciferase gene complex (LUX A; LUX B) from Vibrio Harvey
  • the LUX-gene complex is 3 ', ie "behind" promoter. J the P Loo defined and is therefore transcribed therefrom; the P R_M ..- pro- For its part, motor is negatively controlled by the gene product of the C gene - the lambda repressor; Although this lambda repressor is always synthesized in sufficient quantity by constitutive expression of the P_-dependent C ⁇ gene, it is heat-labile by using the C Ig57 allele.
  • the P_ M promoter remains blocked by binding the repressor until the repressor protein is denatured by increasing the temperature to 37 ° to 42 ° C for 5 to 15 minutes and the P _-.- promotor falls off.
  • pGL3 The DNA-dependent RNA polymerase then binds to the free promoter and causes the massive transcription of the LUX gene complex. Overall, this construction of pGL3 leads to an adjustable, since temperature-dependent synthesis of the enzyme luciferase, which is responsible for the bioluminescence of the indicator organisms.
  • the construction base of pGL3 is the plasmid pLK915 (see KK Stanley, and JP Luzio, EMBO J. 3: 1429 to 1434, 1984).
  • Compatibility of the Sal 1 interface with Bam Hl was determined by previously attaching the 45 base pair long Bam Hl - Sal 1 restriction fragment from the polylinker region of the plasmid "pBluescript" (company "STRATAGENE”; San Diego, Cal., USA ) at the Sal 1 end and subsequent restriction cleavage with the enzyme Bam HI.
  • the above-described plasmid pGL3 can in principle be introduced into any desired species of microorganism by standard techniques in order finally to be cultured in large quantities. It is possible to store bacteria as glycerol cultures at -20 ° C or after freeze-drying at ambient temperature. Freeze-drying is carried out either in glass ampoules or in specially made plastic containers, e.g. in two separate chambers the lyophilized microorganisms or the components necessary for the revitalization of the bacteria. Contain nutrient medium. In order to carry out measurements of toxic substances in a gas phase, the indicator organisms are permanently separated from a measuring compartment by a membrane impermeable to water vapor. In this way, the constant composition of the growth medium and thus the measurement over a longer measurement period, as required for toxicity tests of gases, is ensured.
  • the application of the lyophilized indicator organisms to a suitable carrier material in the form of a test strip is a preferred form of application.
  • the microorganisms are applied to the carrier matrix in a defined amount, fixed by a special process, freeze-dried and finally sealed as a test strip.
  • the indicator bacteria are incubated for one to two hours in aqueous nutrient medium Ambient temperature revitalized.
  • the measuring principle of the toxicity test described here is based on the use of a two-step method:
  • a "screening" is carried out with a number of different test organisms of defined sensitivity for the detection of general toxicity in a sample. Thereafter, further detection reactions can be carried out with strains which are provided with a specific, genetically present or introduced by transformation property of resistance or (hyper) sensitivity.
  • a toxin to which a strain is specifically resistant will lead to an un or only partially reduced bioluminescence signal; in contrast, a non-resistant control strain is more serious in its viability and, correlated with it, its ability to bioluminescence.
  • the measuring method according to the invention is therefore suitable on the one hand for determining the degree of generally present toxicity of a sample through the growth behavior of microorganisms, and on the other hand it is also possible to use all substances with which a specific reactivity of certain test agents ganisms exists to identify and determine their concentration approximately easily and quickly.
  • indicator organisms - this time endowed with specific resistance or sensitivity to certain toxic substances - are incubated with the sample liquid in order to finally carry out bioluminescence measurements. If the bioluminescence signal is unchanged only in one of the test organisms, for example at the level of the control value, " while all other microorganisms experience a reduction in measurable bioluminescence, the toxin to which the strain in question is resistant is identified.
  • a strain of the gram-negative bacterium E. coli ATCC 25922 was treated with a plasmid [(pGL3.T, which in addition to the grain components of pGL3 (see above) contains a tetracycline resistance gene (strain "T” in Fig. 2)].
  • pGL3.T which in addition to the grain components of pGL3 (see above) contains a tetracycline resistance gene (strain "T” in Fig. 2)].
  • strain "C" a derivative of ATCC 25922 - equipped with pGL3 - was used.
  • Several 0.5 ml samples of raw milk with different concentrations of the antibiotic tetracycline were mixed with 0.1 ml (about 2 x 10 bacteria) suspensions of the strains C and T and incubated for 20 minutes at room temperature. After heating the cultures at 40 ° C.
  • FIG. 1 shows the measurement results of samples with five different antibiotic concentrations as relative light signals (RLU; ordinate) as a function of time.
  • the bioluminescence signal of the tetracycline-resistant strain T is relatively constant over the measurement period, while strain C shows a decrease in the amount of light emitted in a concentration-dependent manner.
  • strain C shows a decrease in the amount of light emitted in a concentration-dependent manner.
  • the fact that the growth properties of the indicator strain T are not significantly impaired by tetracycline makes it possible to use this antibiotic in samples (e.g. milk) of unknown origin.
  • the determination of absolute concentrations of this antibiotic or other toxic substances against which an indicator strain is specifically resistant is possible to a good approximation by including reference values of known concentration of the toxin in the measurement series.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Organic Chemistry (AREA)
  • Metallurgy (AREA)
  • Materials Engineering (AREA)
  • General Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • General Engineering & Computer Science (AREA)
  • High Energy & Nuclear Physics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Cleaning In General (AREA)

Abstract

Un dispositif d'électropolissage de surfaces comprend un logement (60) fermé de tous les côtés, à l'exception du côté qui fait face à la surface à polir électriquement. Le logement peut être déplacé le long de la surface à polir (48) par un dispositif de guidage (4, 12, 14, 28). Une garniture d'étanchéité (62) est agencée entre le bord du logement et la surface à polir. Une électrode (56) est assujettie au logement et des accessoires (41, 42) d'alimentation en courant et en un électrolyte fluide sont raccordés au logement. Pour réduire autant que possible les résidus d'électrolyte sur la surface polie, le logement (60) est monté avec l'électrode (56) à l'intérieur d'un logement extérieur (34) fermé de tous les côtés, à l'exception du côté qui fait face à la surface à polir. Le logement extérieur comprend sur son bord une garniture d'étanchéité (50) et il est pourvu de raccords (86, 88) d'amenée et d'évacuation d'un fluide de rinçage, ainsi que de passages pour l'amenée de l'électrolyte fluide jusqu'au logement intérieur (60) et pour l'évacuation de l'électrolyte fluide du logement intérieur (60). Les résidus d'électrolyte qui restent sur des parties des surfaces polies sont ainsi emportés directement derrière le logement intérieur. Le dispositif décrit est particulièrement utile pour polir électriquement les parois internes de récipients, de canalisations et d'autres parties d'installations des industries chimique et nucléaire.
PCT/DE1988/000626 1988-10-10 1988-10-10 Dispositif d'electropolissage de surfaces WO1990004053A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
DE8888908666T DE3878697D1 (de) 1988-10-10 1988-10-10 Vorrichtung zum elektropolieren von oberflaechen.
PCT/DE1988/000626 WO1990004053A1 (fr) 1988-10-10 1988-10-10 Dispositif d'electropolissage de surfaces
EP88908666A EP0436528B1 (fr) 1988-10-10 1988-10-10 Dispositif d'electropolissage de surfaces
US07/683,261 US5135632A (en) 1988-10-10 1991-04-10 Apparatus for electropolishing surfaces

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/DE1988/000626 WO1990004053A1 (fr) 1988-10-10 1988-10-10 Dispositif d'electropolissage de surfaces

Publications (1)

Publication Number Publication Date
WO1990004053A1 true WO1990004053A1 (fr) 1990-04-19

Family

ID=6819755

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1988/000626 WO1990004053A1 (fr) 1988-10-10 1988-10-10 Dispositif d'electropolissage de surfaces

Country Status (3)

Country Link
EP (1) EP0436528B1 (fr)
DE (1) DE3878697D1 (fr)
WO (1) WO1990004053A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998022953A1 (fr) * 1996-11-15 1998-05-28 Babcock Rosyth Defence Limited Limitation de la contamination de la surface d'un element metallique par des radionucleides
US6340572B1 (en) 1991-04-04 2002-01-22 Board Of Regents, The University Of Texas System Kit for the isolation, identification and quantitation of toxicants

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IT1311147B1 (it) * 1999-11-04 2002-03-04 Edk Res Ag Macchina per pulizia localizzata con cella, elettrolitica e/o adultrasuoni, di decapaggio e/o lucidatura

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0146833A2 (fr) * 1983-12-14 1985-07-03 Siemens Aktiengesellschaft Dispositif pour le polissage électrolytique des surfaces intérieures d'objets cylindriques creux

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3370175A (en) * 1965-01-28 1968-02-20 North American Rockwell Toxicant detector
CA1103050A (fr) * 1977-09-28 1981-06-16 Anthony A. Bulich Methode de detection de substances toxiques dans un liquide
US4581335A (en) * 1982-12-01 1986-04-08 Texas A&M University System Process for producing a cloned luciferase-synthesizing microorganism
DE3789766T2 (de) * 1986-07-22 1994-09-29 Texas A & M University Syst Verwendung von bakteriellen luciferase strukturellen genen zum klonieren und zur steuerung der genexpression in mikroorganismen, sowie zur etikettierung und identifizierung von genetisch gebildeten organismen.

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0146833A2 (fr) * 1983-12-14 1985-07-03 Siemens Aktiengesellschaft Dispositif pour le polissage électrolytique des surfaces intérieures d'objets cylindriques creux

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6340572B1 (en) 1991-04-04 2002-01-22 Board Of Regents, The University Of Texas System Kit for the isolation, identification and quantitation of toxicants
WO1998022953A1 (fr) * 1996-11-15 1998-05-28 Babcock Rosyth Defence Limited Limitation de la contamination de la surface d'un element metallique par des radionucleides

Also Published As

Publication number Publication date
EP0436528B1 (fr) 1993-02-24
EP0436528A1 (fr) 1991-07-17
DE3878697D1 (de) 1993-04-01

Similar Documents

Publication Publication Date Title
WO1990004037A1 (fr) PROCEDE POUR DECELER ET IDENTIFIER DES SUBSTANCES TOXIQUES A l'AIDE DE MICROORGANISMES CLONES
Bååth Measurement of heavy metal tolerance of soil bacteria using thymidine incorporation into bacteria extracted after homogenization-centrifugation
DE69310207T2 (de) Methode zum Nachweis von unweltbedingten Vergehen
DE69225221T2 (de) Fusionierte Gene sowie deren Verwendungzur Bestimmung von Metall- und Xenobiotikagehalten
DE69527850T2 (de) Bioluminiscente lyophilisierte bakterien-reagenz zur giftstoff nachweit
DE69734204T2 (de) Lichtproduzierender biosensor
DE3789766T2 (de) Verwendung von bakteriellen luciferase strukturellen genen zum klonieren und zur steuerung der genexpression in mikroorganismen, sowie zur etikettierung und identifizierung von genetisch gebildeten organismen.
AU2017393714B2 (en) Genotoxic substance detection vector and detection method thereof
DE69121888T2 (de) Screening nach proteinpartnern und deren verwendung
EP0456667B1 (fr) Procede de depistage du mercure au moyen de micro-organismes dont la bioluminescence augmente en presence de mercure
WO1990004053A1 (fr) Dispositif d'electropolissage de surfaces
Wingfield et al. The effect of soil treatment on the response of the soil microflora to the herbicide dalapon
DE4343527A1 (de) Verfahren zur Identifizierung von Stoffen mit potentieller herbizider oder wachstumsregulatorischer Wirkung mittels pflanzlicher Transporterproteine, Verwendung der Transporterproteine sowie Substanzen mit herbizider und wachstumsregulatorischer Wirkung
Rolfe et al. Density differences between genetic markers in Pneumococcal transforming principle
DE102007027619A1 (de) Biosensor
DE69934847T2 (de) Diagnosesystem und verfahren zur bestimmung des vorhandenseins einer genotoxischen verbindung in einer probe
DE69826029T2 (de) Acetyl-coa-carboxylase von candida albicans
DE69231710T2 (de) Tyrosinkinase
DE10061872A1 (de) Hefestamm zur Prüfung der Geno- und Zytotoxizität komplexer Umweltkontaminationen
EP0143107B1 (fr) Méthode pour la détection de virus dans des cultures pures d'organismes microscopiques ainsi que dans des comestibles et dans des produits technologiques
DE19544374C1 (de) Plasmid und damit transformierter Mikroorganismus für ein Verfahren zur luminometrischen Bestimmung der mutagenen und/oder karzinogenen Potenz eines Agens
EP1199560B1 (fr) Procédé électrochimique pour la détermination du nombre de cellules et l'activité des systèmes biologiques
DE69838175T2 (de) Plasmide aus ammonium oxidierenden bakterien und ihre verwendung
EP1402057A2 (fr) Methode de detection de substances mutagenes
DE112007001823T5 (de) Prüfung der Wirksamkeit von Konservierungsmitteln

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE FR GB IT LU NL SE

CFP Corrected version of a pamphlet front page
CR1 Correction of entry in section i

Free format text: IN PAT.BUL.09/90,THE ERRONEOUSLY PUBLISHED FIGURE REPLACED BY THE CORRECT FIGURE

WWE Wipo information: entry into national phase

Ref document number: 1988908666

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1988908666

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 1988908666

Country of ref document: EP

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载