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21 results about "Origin of replication" patented technology
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The origin of replication (also called the replication origin) is a particular sequence in a genome at which replication is initiated. Propagation of the genetic material between generations requires timely and accurate duplication of DNA by semiconservative replication prior to cell division to ensure each daughter cell receives the full complement of chromosomes. This can either involve the replication of DNA in living organisms such as prokaryotes and eukaryotes, or that of DNA or RNA in viruses, such as double-stranded RNA viruses. Synthesis of daughter strands starts at discrete sites, termed replication origins, and proceeds in a bidirectional manner until all genomic DNA is replicated. Despite the fundamental nature of these events, organisms have evolved surprisingly divergent strategies that control replication onset. Although the specific replication origin organization structure and recognition varies from species to species, some common characteristics are shared.
Large vectors and methods for high-yield production
Provided herein are methods for the production of a vector with a size of at least 16 kb from bacterial cells comprising the consecutive steps of a) obtaining bacterial cells comprising a vector with a size of at least 16 kb, comprising an inducible origin of replication, b) inoculating culture medium with the bacterial cells comprising the vector, c) culturing the bacterial cells in the culture medium, d) adding one or more inducers of said inducible origin of replication to the culture medium when the bacterial culture has reached an optical density at 600 nm (OD600) of at least 20, e) further culturing the bacterial cells in the culture medium, f) optionally separating the bacterial cells from the culture medium, and g) recovering the plasmid from the bacterial cells. Also provided herein are vectors with a size of at least 16 kb comprising an inducible origin of replication for use in such methods.
Owner:KATHOLIEKE UNIV LEUVEN
A specific targeted method to eliminate bladder without leaving scars. KPC-2 Gene and IncL plasmid sgRNA sequences, CRISPR / Cas9 system and applications
This invention belongs to the field of bioengineering technology and discloses a method for specific targeting and scarless simultaneous elimination blue KPC‑2 Gene and IncL plasmid sgRNA sequences, CRISPR / Cas9 vector system and applications. This system targets the sgRNA of the IncL plasmid. blue KPC‑2 The IncL-type plasmid of the gene was designed with an sgRNA that specifically recognizes the conserved sequence at its origin of replication, thus eliminating the drug resistance gene while disrupting the plasmid's replication ability. Conjugation transfer to recipient bacteria significantly improved vector delivery efficiency. An arabinose-inducible promoter was used to control Cas9 gene expression, improving system stability and editing success rate. The vector carries a sucrose-inducible self-eliminating gene, enabling controlled vector self-elimination. This system is not a simple superposition but rather constitutes a time-controlled operational flow: efficient delivery via conjugation, controlled cleavage via induction, and finally, induced self-elimination.
Owner:KUNMING UNIV OF SCI & TECH
Improved generation of viral and nonviral nanoplasmid vectors
PendingJP2026116424AOrigin of replicationInverted Repeat Sequences
A method for improving the replication of covalent closed circular plasmids is provided. [Solution] The method comprises the step of preparing a covalent closed circular plasmid having a Pol I-dependent origin of replication and an insert containing a structured DNA sequence selected from the group consisting of a reverse repeat sequence, a directional repeat sequence, a homopolymer repeat sequence, a eukaryotic origin of replication, or a eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also comprises the step of modifying the covalent closed circular recombinant molecule so that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, thereby improving the replication of the covalent closed circular plasmid with the resulting Pol III-dependent origin of replication. A covalent closed circular recombinant DNA molecule without antibiotic markers is also provided.
Owner:ALDEVRON LLC
Optimized lentiviral transfer vectors and uses thereof
PendingUS20260062712A1Polypeptide with localisation/targeting motifImmunoglobulin superfamilyHeterologousOrigin of replication
The invention features lentiviral transfer vectors that include heterologous nucleic acids to be introduced into a cell. The lentiviral transfer vector may be characterized by the following features: (a) including a cytomegalovirus (CMV) promoter; (b) including a polynucleotide encoding a partial gag protein that includes a mutated INS1 inhibitory sequence that reduces restriction of nuclear export of RNA; (c) not including a polynucleotide encoding the INS2, INS3, and INS4 inhibitory sequences of gag; (d) not including an SV40 origin of replication and / or an f1 origin of replication; (e) including a cPPT sequence that contains splice site; (f) including an EF1alpha promoter with intact splice donor and acceptor sites; and (g) including hepatitis B PRE with mutation in start codon of X protein ORF.
Owner:NOVARTIS AG +1
CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / CRISPR / Cas9) system for specific targeted traceless elimination of blaNDM-1 gene and IncN plasmid and application
PendingCN121950810AOvercome the bottleneck of conversion difficultiesImprove delivery efficiencyHydrolasesMicroorganism based processesGene deliveryOrigin of replication
The invention belongs to the technical field of biological genetic engineering, and discloses a CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / associated protein 9) system for specific targeted traceless elimination of a blaNDM-1 gene and an IncN plasmid and application of the CRISPR / Cas9 system. According to the system, sgRNA of an original point conserved sequence is copied by designing an IncN type plasmid carrying a blaNDM-1 gene in a targeting manner, so that the replication capability of the plasmid is destroyed while a drug-resistant gene is eliminated; the carrier delivery efficiency is greatly improved by conjugational transfer to recipient bacteria; the expression of the Cas9 gene is controlled by adopting an arabinose inducible promoter, so that the constitutive toxicity of Cas9 protein is reduced, and the system stability and the editing success rate are improved; the sucrose-induced carrier self-elimination gene carried by the carrier realizes controllable self-elimination of the carrier. The system is not simply superposed, but forms an operation process with a controllable time sequence: efficient delivery is firstly carried out through joint, then cutting is controlled through induction, and finally self-elimination is induced.
Owner:KUNMING UNIV OF SCI & TECH
DNA production methods
PCT designated stageWO2026102108A1Genetic material ingredientsFermentationOrigin of replicationDouble strand
The disclosure provides, for example, a method of making a composition comprising purified DNA molecules. In some embodiments, the method comprises contacting a solution comprising a plurality of circular, double-stranded DNA molecules to a hydrophobic interaction chromatography matrix. In some embodiments, the circular, double-stranded DNA molecules lack one or both of a bacterial origin of replication and a selectable marker.
Owner:FLAGSHIP PIONEERING INNOVATIONS VII LLC
Proteus mirabilis homologous recombination system expression plasmid, expression system, application and gene knockout / knockin method
PendingCN122012561AMicroorganism based processesStable introduction of DNAHeterologousOrigin of replication
The invention belongs to the field of biology, and discloses a proteus mirabilis homologous recombination system expression plasmid which comprises a pBR322 replication starting point, a resistance gene, an arabinose inducible promoter and a proteus mirabilis-derived homologous recombination operon. The homologous recombination operon is used for coding a TPM66325 protein, an EPM66325 protein, a single-chain binding protein SSB and a fake release protein. The expression plasmid has relatively high recombination efficiency in proteus mirabilis GDMCC 66325 and can also play a recombination role in a proteus mirabilis standard strain ATCC35659, and the expression plasmid is obtained by knocking out a related gene bcsB formed by a hemolysin related gene hpmA and a biological membrane of the proteus mirabilis GDMCC 66325 and directly knocking in a heterologous gene after a related gene cluster is adjusted by flagellum. The effectiveness of the expression plasmid in recombinant operation of proteus mirabilis is verified; meanwhile, the invention further discloses a proteus mirabilis homologous recombination system, application and a gene knockout / knockin method.
Owner:SOUTH CHINA AGRICULTURAL UNIVERSITY
Biosynthesis method
The present invention relates generally to methods for producing products by cells, in particular to the field of industrial biotechnology or microbiology, in which useful chemicals and products, such as biological products such as proteins, are produced using living cells. The present invention relates to a method of producing a product by a cell, the method comprising: excising one or more nucleic acid sequences from a chromosome in the cell wherein the one or more nucleic acid sequences comprise a gene of interest and a starting point of replication; forming a DNA molecule outside the chromosome, wherein the DNA molecule outside the chromosome comprises a resected target gene and a replication starting point which are resected; and inducing expression of the target gene from the DNA molecule outside the chromosome. The invention also relates to cells for producing the product, polynucleotide vectors and uses thereof.
Owner:GEARBOX BIOSCIENCES EUROPE
Mutant vectors for improved host transformation
PCT designated stageWO2025255554A1Microbiological testing/measurementVirus peptidesOrigin of replicationReplication initiator protein
The invention provides methods of screening replication initiator protein (Rep) mutants that modulate vector's copy number of different origins of replication within Agrobacterium. The invention also provides compositions comprising such Rep mutants that can improve Agrobacterium-mediated transformation efficiency of a vector, such as a binary vector, to a host cell.
Owner:RGT UNIV OF CALIFORNIA
Viral and non-viral nanoplasmid vectors with improved production
ActiveUS12600984B2Microorganism based processesNucleic acid vectorOrigin of replicationInverted Repeat Sequences
A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I-dependent origin of replication, and an insert including a structured DNA sequence selected from inverted repeat sequences, direct repeat sequences, homopolymeric repeat sequences, eukaryotic origins of replication or eukaryotic promoter enhancer sequences, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also includes modifying the covalently closed circular recombinant molecule such that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, whereby the resultant Pol III-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinant DNA molecule is also provided.
Owner:ALDEVRON LLC
Biosynthetic methods
The present invention generally relates to methods for producing products using cells, and more particularly to the field of industrial biotechnology or industrial microbiology in which living cells are used to produce useful chemical substances and products, such as biological products such as proteins. The present invention relates to a method for producing products using cells, comprising the steps of: excising one or more nucleic acid sequences including a target gene and an origin of replication from a chromosome within a cell; forming an extrachromosomal DNA molecule including the excised target gene and the excised origin of replication; and inducing the expression of the target gene from the extrachromosomal DNA molecule. The present invention also relates to cells, polynucleotide vectors, and their use for producing products.
Owner:オウ ギアボックス バイオサイエンシーズ
Virus-based expression of proteins and genes
PCT designated stageWO2026175928A1Origin of replicationMeasles virus IgG
A nucleic acid molecule in the form of a plasmid comprising an origin of replication, a promoter for the transcription into viral RNA; a 3'NCT triplet upstream of regions for the insertion of gene(s) of interest, separated by Measles virus intergenic regions, and one 5'NCT sharing at least 95% of identity with SEQ ID NO: 11 over its full-length, wherein the 3'NCT triplet shares at least 95% of identity with SEQ ID NO:14 over its full-length, the corresponding helper nucleic acid molecules, a process for the production of virosomes based on these nucleic acid based molecules, the obtained virosomes and their therapeutical uses.
Owner:SAGITTA BIOTECH
Baculovirus expression systems and methods thereof
PCT designated stageWO2026013698A3Virus peptidesStable introduction of DNAHeterologousOrigin of replication
An improved baculovirus expression system for enhanced heterologous protein expression in hosts selected from Bombyx mori larvae, pupae, primary cells, or established insect cell lines is disclosed. The expression system is derived from a wild-type strain of Bombyx mori nucleopolyhedrovirus (BmNPV) by deletion or inactivation of non-essential genes, of the BmNPV genome. The system includes novel transfer vectors designed for targeted gene modification and incorporation of a bacterial origin of replication and an insertion site for a gene of interest (GOI). This engineered bacmid maintains infectivity while offering enhanced genetic stability and significantly improved heterologous protein expression in insect cells, larvae, or pupae. Methods for constructing this bacmid via homologous recombination and using it for high-level protein production are also described.
Owner:LOOPWORM PVT LTD
Plasmid vectors, shuttle vectors, and protein manufacturing methods
To provide plasmid DNA with a novel replication origin that can stably introduce foreign genes into actinomycetes. [Solution] The plasmid vector according to the present disclosure comprises (a) a nucleotide sequence having a specific sequence, or (b) a nucleotide sequence in which one or more nucleotides are substituted, deleted and / or inserted in the nucleotide sequence having the specific sequence.
Owner:IWATE UNIVERSITY
Gene Editing System of Escherichia Coli and Gene Editing Method Thereof
A gene editing system of Escherichia coli includes an Escherichia coli, a helper plasmid and a donor plasmid. The helper plasmid successively includes a transposase complex expression cassette, a Cas12k expression cassette, a first sgRNA cassette, a first antibiotic resistance gene and a first replication origin. The donor plasmid successively includes a left end sequence of a ShCAST transposon, an exogenous gene expression cassette, a right end sequence of the ShCAST transposon, a second sgRNA cassette, a second antibiotic resistance gene and a second replication origin.
Owner:NATIONAL TSING HUA UNIVERSITY
Plasmids with an increased replication rate
PCT designated stageWO2025252302A1Nucleic acid vectorVector-based foreign material introductionOrigin of replicationPlasmid Vector
The invention relates to a plasmid vector comprising a) an origin of replication; and b) a PAS region, comprising i) a primosomal assembly sequence PAS-forward (SEQ ID NO: 2), which is located 3´ of the origin of replication and in the direction of replication; ii) a primosomal assembly sequence PAS-reverse (SEQ ID NO: 3), which is located 3´ of the primosomal assembly sequence PAS-forward and in the direction opposite the replication; and iii) a sequence X between the primosomal assembly sequence PAS-forward and the primosomal assembly sequence PAS-reverse, said sequence sharing a sequence identity of at most approximately 75% with SEQ ID NO: 1.
Owner:WACKER CHEMIE AG
Baculovirus expression systems and methods thereof
PCT designated stageWO2026013698A2Virus peptidesStable introduction of DNAHeterologousOrigin of replication
An improved baculovirus expression system for enhanced heterologous protein expression in hosts selected from Bombyx mori larvae, pupae, primary cells, or established insect cell lines is disclosed. The expression system is derived from a wild-type strain of Bombyx mori nucleopolyhedrovirus (BmNPV) by deletion or inactivation of non-essential genes, of the BmNPV genome. The system includes novel transfer vectors designed for targeted gene modification and incorporation of a bacterial origin of replication and an insertion site for a gene of interest (GOI). This engineered bacmid maintains infectivity while offering enhanced genetic stability and significantly improved heterologous protein expression in insect cells, larvae, or pupae. Methods for constructing this bacmid via homologous recombination and using it for high-level protein production are also described.
Owner:LOOPWORM PVT LTD
Cloning / expression vectors for non-B-type DNA sequences
Provided is a vector comprising i) an origin of replication, ii) a first terminator, iii) a) a target DNA sequence comprising a DNA sequence capable of forming a non-B-type DNA configuration, or b) a target DNA expression cassette comprising a promoter and a target DNA sequence wherein the target DNA sequence comprises a DNA sequence capable of forming a non-B-type DNA configuration, iv) a second terminator, and v) a selection marker and a promoter thereof, wherein the replication starting point is at least 150bp apart from the DNA sequence capable of forming the non-B-type DNA configuration in the replication direction, and the first terminator and the second terminator are respectively located at the upstream and downstream of the target DNA sequence of a), or the first terminator and the second terminator are respectively located at the upstream and downstream of the target DNA expression cassette of b). The provided vector can improve the amplification and / or expression (transcription) stability of the poly (A) tail and the ITR sequence.
Owner:NANJING GENSCRIPT BIOTECH CO LTD
Expression constructs, cells and methods of obtaining cells
To provide an expression construct capable of improving resistance to a specific target substance, a cell transformed by the expression construct, and a method for obtaining a cell by the expression construct.SOLUTION: A first promoter having constitutive activity and functioning in a cell, a first nucleotide sequence encoding a CRISPR enzyme operably linked to the first promoter, a second promoter functioning in the cell, a second nucleotide sequence encoding a guide RNA operably linked to the second promoter, a third promoter for expression and functioning in the cell, a third nucleotide sequence encoding a protein for introducing a point mutation into the genome of the cell operably linked to the third promoter, a drug resistance gene, and a replication origin, wherein the guide RNA is capable of complementarily binding to at least a part of a multicopy sequence present in the genome.SELECTED DRAWING: Figure 1
Owner:PUBLIC UNIVERSITY CORPORATION OSAKA CITY UNIVERSITY
Klebsiella pneumoniae homologous recombination system and application thereof
The invention belongs to the field of microbial genetic engineering, and relates to a homologous recombination system in gram-negative bacteria, in particular to a klebsiella pneumoniae homologous recombination system and application thereof. Comprising a pBBR1 replication starting point, a kanamycin resistance gene, a rhamnose inducible promoter and a klebsiella pneumoniae source homologous recombination operon or a mutant of the klebsiella pneumoniae source homologous recombination operon. According to the invention, self homologous recombinant protein of klebsiella pneumoniae is excavated, and expression plasmids of the homologous recombination system of klebsiella pneumoniae are constructed, so that the application of the homologous recombination system of klebsiella pneumoniae in genome DNA genetic modification of short homologous arm mediated homologous recombination in klebsiella pneumoniae and other klebsiella pneumoniae is realized; or gene knockout and insertion are carried out on a bacteriophage genome.
Owner:SHANDONG UNIV
Constitutive expression plasmid vector for high expression of xanthine oxidase and application of constitutive expression plasmid vector
The invention belongs to the technical field of gene engineering, and discloses a constitutive expression plasmid vector for high expression of xanthine oxidase and application of the constitutive expression plasmid vector. The plasmid vector, as a heterologous expression system, can be used for efficiently expressing the recombinant xanthine oxidase derived from the cellulomicrobe TH20 in Pseudomonas putida KT2440. Specifically, a replication starting point of pUCP18 is replaced by a replication starting point of pSC101 with a low copy number, and a constitutive expression plasmid pPegP119 is constructed, so that the metabolic burden is relieved. A TH20 XOD gene cluster is cloned into the vector to construct a recombinant strain, and xanthine oxidase can be efficiently produced at low cost. According to the invention, the high-efficiency expression of the xanthine oxidase is realized in Pseudomonas putida KT2440 through a gene recombination technology, and the maximum expression quantity of the xanthine oxidase can be realized without adding an inducer in the expression.
Owner:BEIJING INSTITUTE OF PETROCHEMICAL TECHNOLOGY