WO1996012015A1 - Boucle d'arn/adn auto-stabilisante - Google Patents

Boucle d'arn/adn auto-stabilisante Download PDF

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Publication number
WO1996012015A1
WO1996012015A1 PCT/NL1995/000344 NL9500344W WO9612015A1 WO 1996012015 A1 WO1996012015 A1 WO 1996012015A1 NL 9500344 W NL9500344 W NL 9500344W WO 9612015 A1 WO9612015 A1 WO 9612015A1
Authority
WO
WIPO (PCT)
Prior art keywords
dna
gene
binding protein
rna
host
Prior art date
Application number
PCT/NL1995/000344
Other languages
English (en)
Inventor
Evert Hovius
Original Assignee
Evert Hovius
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 Evert Hovius filed Critical Evert Hovius
Priority to AU37550/95A priority Critical patent/AU3755095A/en
Publication of WO1996012015A1 publication Critical patent/WO1996012015A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli

Definitions

  • the SSL is formed by incorporating a DNA/RNA strand
  • prokaryotes or eukaryotes whereby a loop is formed consisting of a part of the DNA introduced and the inefficient operon or gene.
  • the loop is stabilized by a binding protein (to be examined further), the gene of which is located on the DNA strand introduced.
  • the positive regulator of the binding protein is located before the gene thereof.
  • the negative regulator inter alia together with the inefficient operon or gene, is excluded in the loop.
  • Located behind the gene of the binding protein are, respectively, a leader sequence, attenuator site, recognition sequence for the binding protein, negative regulator, attenuator site, leader sequence followed by a series of initiation signals.
  • the DNA/RNA strand is so composed as to specifically nestle before the inefficient operon or gene (or whatever it is desired to exclude in the loop), in that the stability of the total DNA is increased.
  • the manipulated DNA/RNA is situated in a protein coat the code of which does not occur on the DNA/RNA, as a result it cannot reproduce.
  • the concentration thereof must be as high as possible.
  • the lactose operon is mutated in one of the structural genes, then the proper form of the operon is incorporated into a manipulated virus (DNA/RNA strand) followed by a positive regulator gene A1.
  • the RNA polymerase starts the transcription on the lactose operon on the virus DNA. Thereafter the positive regulator gene A1 is transcribed, followed by the transcription of the binding gene B.
  • the binding gene B contains no normal termination site, but is followed by a leader sequence which contains two fantasy codons. As a result, the ribosomes stall, so that the leader messenger RNA passes a signal to the RNA polymerase to stop the transcription.
  • the RNA polymerase reaches the attenuator site and leaves the DNA here. Because the RNA polymerase leaves the DNA here, it does not reach the loop linked by the binding protein B.
  • RNA polymerase scans the DNA for
  • L leader sequence
  • the RNA polymerase falls off the virus DNA. (slide effect).
  • the attenuator site is followed by the
  • binding protein B binding protein B between two attenuator sites.

Landscapes

  • Genetics & Genomics (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Il s'agit d'un groupe de nouveaux médicaments d'origine génétique susceptibles de se substituer aux antibiotiques actuels, et constitués d'un ADN/ARN exempt de gène d'enveloppe et manipulé de telle manière qu'il se fixe dans une position spécifique et que, entre autres, il contient, dans l'ordre, les séquences suivantes: 1) l'opéron ou gène éventuellement à substituer; 2) un régulateur positif de la protéine de liaison; 3) le gène pour la protéine de liaison; 4) une séquence initiale; 5) un site atténuateur; 6) une séquence de reconnaissance pour la protéine de liaison; 7) un régulateur négatif pour la protéine de liaison; 8) un site atténuateur; 9) une séquence initiale; et 10) une séquence riche en initiation. Par conséquent, une boucle se forme dans l'ADN de l'hôte, cette boucle étant constituée d'une partie de l'ADN viral, à savoir les points 6, 7, 8, 9, 10, ainsi que l'opéron ou gène de l'hôte. L'ADN/ARN manipulé se situe dans une enveloppe protéique naturelle ou synthétique qui peut, suivant l'objectif envisagé, reconnaître un hôte.
PCT/NL1995/000344 1994-10-12 1995-10-10 Boucle d'arn/adn auto-stabilisante WO1996012015A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU37550/95A AU3755095A (en) 1994-10-12 1995-10-10 Self stabilizing rna/dna loop

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
NL9401678A NL9401678A (nl) 1994-10-12 1994-10-12 Methode van de zichzelf stabiliserende lus (Z.Z.S.L.).
NL9401678 1994-10-12

Publications (1)

Publication Number Publication Date
WO1996012015A1 true WO1996012015A1 (fr) 1996-04-25

Family

ID=19864761

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/NL1995/000344 WO1996012015A1 (fr) 1994-10-12 1995-10-10 Boucle d'arn/adn auto-stabilisante

Country Status (3)

Country Link
AU (1) AU3755095A (fr)
NL (1) NL9401678A (fr)
WO (1) WO1996012015A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2511033A1 (fr) * 1981-08-10 1983-02-11 Genex Corp Vecteurs d'expression permettant d'introduire un gene dans un organisme procaryote
WO1990000192A1 (fr) * 1988-07-01 1990-01-11 Genencor, Inc. Champignons filamenteux a carence de proteinase aspartique

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2511033A1 (fr) * 1981-08-10 1983-02-11 Genex Corp Vecteurs d'expression permettant d'introduire un gene dans un organisme procaryote
WO1990000192A1 (fr) * 1988-07-01 1990-01-11 Genencor, Inc. Champignons filamenteux a carence de proteinase aspartique

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
AMOUYAL EN VON WILCKEN-BERGMANN: "Repression of the E.coli lactose operon by cooperation between two individually unproductive "half-operator" sites", COMPTES RENDUS HEBDOMADAIRES DES SEANCES DE L'ACADEMIE DES SCIENCES, SERIE C: SCIENCES CHIMIQUES, vol. 315, MONTREUIL FR, pages 403 - 407 *

Also Published As

Publication number Publication date
AU3755095A (en) 1996-05-06
NL9401678A (nl) 1996-05-01

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