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1. (WO2019041058) GENE CONSTRUCT AND BIOSENSOR FOR THE RAPID DETECTION OF AHL MOLECULES AND THE PATHOGENIC BACTERIA THAT PRODUCE SAME
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Pub. No.: WO/2019/041058 International Application No.: PCT/CL2018/050077
Publication Date: 07.03.2019 International Filing Date: 30.08.2018
IPC:
C12N 1/21 (2006.01) ,C12N 15/63 (2006.01) ,C12N 15/70 (2006.01) ,C12Q 1/04 (2006.01) ,C12R 1/19 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1
Micro-organisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving micro-organisms or compositions thereof; Processes of preparing or isolating a composition containing a micro-organism; Culture media therefor
20
Bacteria; Culture media therefor
21
modified by introduction of foreign genetic material
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70
Vectors or expression systems specially adapted for E. coli
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
02
involving viable micro-organisms
04
Determining presence or kind of micro-organism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
R
INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C-C12Q75
1
Micro-organisms
01
Bacteria or actinomycetales
185
Escherichia
19
Escherichia coli
Applicants:
UNIVERSIDAD DE VALPARAÍSO [CL/CL]; Blanco 951 Valparaíso, 2391415, CL
Inventors:
DINAMARCA, Alejandro; CL
ROMO, Natalia; CL
IBACACHE, Claudia; CL
Agent:
JARRY IP SPA; Isidora Goyenechea 3250, of. 302, Las Condes Santiago, 7550083, CL
MORENO SAMPER, Olga; CL
Priority Data:
2201-201730.08.2017CL
Title (EN) GENE CONSTRUCT AND BIOSENSOR FOR THE RAPID DETECTION OF AHL MOLECULES AND THE PATHOGENIC BACTERIA THAT PRODUCE SAME
(FR) CONSTRUCTION GÉNÉTIQUE ET BIOCAPTEUR POUR LA DÉTECTION RAPIDE DE MOLÉCULES AHLS ET LES BACTÉRIES PATHOGÈNES QUI LES PRODUISENT
(ES) CONSTRUCCIÓN GÉNICA Y BIOSENSOR PARA LA RÁPIDA DETECCIÓN DE MOLÉCULAS AHLS Y LAS BACTERIAS PATÓGENAS QUE LAS PRODUCEN
Abstract:
(EN) The invention relates to a gene construct for detecting the presence of microorganisms that produce acyl-homoserine-lactone (AHL) chemical molecules, which gene construct comprises a first expression cassette that includes a copper-inducible promoter operatively bound to the gene that encodes RhiR protein and, downstream of the first expression cassette, a second expression cassette that includes a promoter induced by AHL-RhiR complex, which promoter is operatively bound to a gene that encodes a reporter protein. The invention also relates to a genetically modified biosensor cell for detecting the presence of microorganisms, which comprises said gene construct, and to a method for detecting the presence of microorganisms that produce AHL chemical molecules, by immobilising the biosensor in an organic matrix, thus generating a surface that can be used to display a sample of a fluid such as air or a liquid medium, and thereby detect said molecules using the biosensor cell.
(FR) L'invention concerne une construction génétique de détection de la présence de micro-organismes qui produisent des molécules chimiques du type des homosérine lactones (AHL) qui comprennent une première cassette d'expression qui comprend un promoteur pouvant être induit par du cuivre relié fonctionnel au gène qui codifie la protéine RhIR et, en aval de cette première cassette d'expression, une deuxième cassette d'expression qui comprend un promoteur qui est induit par le complexe AHL-RhIR, ce promoteur étant relié fonctionnel à un gène qui codifie une protéine rapporteuse. L’invention concerne également un cellule biodétectrice modifiée génétiquement pour détecter la présence de micro-organismes qui comprend cette construction génétique, ainsi que le procédé pour détecter la présence de micro-organismes qui produisent des molécules chimiques du type AHL par immobilisation du biocapteur dans une matrice organique, produisant une surface qui peut être utilisée pour exposer un échantillon de fluide tel que l’air ou un milieu liquide, et ainsi effectuer la détection de cette molécule médiée par la cellule de biocapteur.
(ES) La invención se refiere a una construcción génica para detectar la presencia de microorganismos que producen moléculas químicas del tipo de las Acil-homoserina lactonas (AHL) que comprende un primer casete de expresión que incluye un promotor inducible por cobre operativamente unido al gen que codifica la proteína RhlR y, corriente abajo de dicho primer casete de expresión, un segundo casete de expresión que incluye un promotor que se induce por el complejo AHL-RhlR, estando dicho promotor operativamente unido a un gen que codifica una proteína reportera. La invención también incluye una célula biosensora modificada genéticamente para detectar la presencia de microorganismos que comprende dicha construcción génica, así como el método para detectar la presencia de microorganismos que producen moléculas químicas del tipo AHL mediante la inmovilización del biosensor en una matriz orgánica, generando una superficie que se puede utilizar para exponer una muestra de un fluido tal como aire o un medio líquido, y así efectuar la detección de dichas moléculas mediada por la célula biosensora.
Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Spanish (ES)
Filing Language: Spanish (ES)