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1. WO2019128837 - IMPROVED PROMOTER AND CARRIER COMPOSED OF SAME AND APPLICATION THEREOF

Publication Number WO/2019/128837
Publication Date 04.07.2019
International Application No. PCT/CN2018/122312
International Filing Date 20.12.2018
IPC
C12N 15/113 2010.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
113Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides
C12N 15/70 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70Vectors or expression systems specially adapted for E. coli
C12N 15/66 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
66General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
C12N 1/21 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
1Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
20Bacteria; Culture media therefor
21modified by introduction of foreign genetic material
C12R 1/19 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C-C12Q75
1Microorganisms
01Bacteria or actinomycetales
185Escherichia
19Escherichia coli
CPC
C12N 15/66
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
66General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
C12N 15/70
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70Vectors or expression systems specially adapted for E. coli
C12N 2800/22
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
2800Nucleic acids vectors
22Vectors comprising a coding region that has been codon optimised for expression in a respective host
C12N 2800/60
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12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
2800Nucleic acids vectors
60Vectors containing traps for, e.g. exons, promoters
C12N 9/2471
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
14Hydrolases (3)
24acting on glycosyl compounds (3.2)
2402hydrolysing O- and S- glycosyl compounds (3.2.1)
2468acting on beta-galactose-glycoside bonds, e.g. carrageenases (3.2.1.83; 3.2.1.157); beta-agarase (3.2.1.81)
2471Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
C12Y 302/01023
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
YENZYMES
302Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
01Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
01023Beta-galactosidase (3.2.1.23), i.e. exo-(1-->4)-beta-D-galactanase
Applicants
  • 苏州金唯智生物科技有限公司 GENEWIZ INC., SZ [CN]/[CN]
Inventors
  • 薛高旭 XUE, Gaoxu
  • 贾延凯 JIA, Yankai
  • 齐甜铭 QI, Tianming
  • 冯爱华 FENG, Aihua
  • 谢正立 XIE, Zhengli
  • 吴昕 WU, Xin
  • 孙中平 SUN, Zhongping
  • 廖国娟 LIAO, Guojuan
Agents
  • 北京品源专利代理有限公司 BEYOND ATTORNEYS AT LAW
Priority Data
201711490305.530.12.2017CN
Publication Language Chinese (zh)
Filing Language Chinese (ZH)
Designated States
Title
(EN) IMPROVED PROMOTER AND CARRIER COMPOSED OF SAME AND APPLICATION THEREOF
(FR) PROMOTEUR AMÉLIORÉ ET SUPPORT COMPOSÉ DE CELUI-CI ET APPLICATION CORRESPONDANTE
(ZH) 一种改进的启动子及其组成的载体和应用
Abstract
(EN) An improved promoter and application thereof; the improvement refers to the mutation of a nucleotide sequence, in the promoter area, between -35 area to -10 area into an endonuclease recognition site. The improvement may overcome the problem existing in blue-white screening vectors that poisonousness may be produced to the host, which leads to clone incapability, when transcription or translation product of a foreign gene is promoted by a strong promoter. Thus, the defect of false positive clone resulted from gene frame shift mutation which is caused by the deficiency of the vector of 1-2bp at the restriction enzyme cutting site can be avoided, and the false negative phenomenon of a panel being full of locus coeruleus due to smaller foreign DNA fragments and the insertion of foreign DNA not changing the reading frame of the gene can be eliminated.
(FR) L'invention concerne un promoteur amélioré et une application correspondante ; l'amélioration se réfère à la mutation d'une séquence nucléotidique, dans la zone de promoteur, entre la zone -35 et la zone -10 dans un site de reconnaissance d'endonucléase. L'amélioration peut surmonter le problème existant dans des vecteurs de criblage bleu-blanc selon lequel une toxicité pour l'hôte peut être produite, ce qui conduit à l'incapacité de clonage, lorsque le produit de transcription ou de traduction d'un gène étranger est promu par un promoteur fort. Ainsi, le défaut d'un clone faux positif résultant d'une mutation de décalage de cadre génique qui est provoqué par la délétion du vecteur de 1-2 pb au niveau du site de coupe d'enzyme de restriction peut être évité et le phénomène faux négatif d'un panel complet de locus coeruleus dû à des fragments d'ADN étrangers plus petits et l'insertion d'un ADN étranger ne modifiant pas le cadre de lecture du gène peuvent être éliminés.
(ZH) 一种改进的启动子及其应用,该改进为将启动子区域中的-35区至-10区之间的核酸序列突变为核酸内切酶识别位点。能够克服蓝白筛选载体中存在强启动子启动外源基因的转录或翻译产物可能对宿主有毒性而导致无法克隆的问题,可以避免载体在酶切位点缺失1-2bp导致基因移码突变产生假阳性克隆的缺陷,可以消除由于外源DNA片段较小并且外源DNA的插入没有改变基因的读码框而造成的平板都是蓝斑的假阴性现象。
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