174 results about "Chromosomal dna" patented technology

Gene targeting is a technique to introduce genetic change into one or more specific locations in the genome of a cell. For example, gene targeting can introduce genetic change by modifying, repairing, attenuating or inactivating a target gene or other chromosomal DNA. In one aspect, this disclosure relates to methods and compositions for gene targeting with high efficiency in a cell. This disclosure also relates to methods of treating or preventing a genetic disease in an individual in need thereof. Further disclosed are chimeric nucleases and vectors encoding chimeric nucleases.

Chromosomal abnormalities are responsible for a significant number of birth defects, including mental retardation. The present invention is related to methods for non-invasive and rapid, prenatal diagnosis of chromosomal abnormalities based on analysis of a maternal blood sample. The invention exploits the differences in DNA between the mother and fetus, for instance differences in their methylation states, as a means to enrich for fetal DNA in maternal plasma sample. The methods described herein can be used to detect chromosomal DNA deletions and duplications. In a preferred embodiment, the methods are used to diagnose chromosomal aneuploidy and related disorders, such as Down's and Turner's Syndrome.

Synthesis of a target transcript of a gene is selectively increased in a mammalian cell by contacting the cell with a polynucleotide oligomer of 12-28 bases complementary to a region within a target promoter of the gene under conditions whereby the oligomer selectively increases synthesis of the target transcript.

An object of the present invention is to provide a transformation system for Labyrinthulomycota that allows the elucidation of biosynthetic mechanisms of lipids such as PUFA and carotenoids as well as for the construction of a high production system and the design and development of novel functional lipid molecules by the control of the mechanisms. The present invention provides a method for introducing a transgene into a cell of Labyrinthulomycota, which comprises introducing into a cell of Labyrinthulomycota a recombinant vector comprising a transgene and a nucleotide sequence which is homologous to a part of chromosomal DNA of Labyrinthulomycota and is capable of homologous recombination with the chromosomal DNA, and then inducing homologous recombination in this homologous nucleotide sequence.

The invention relates to recombination systems and methods for eliminating nucleic acid sequences from the chromosomal DNA of eukaryotic organisms, and to transgenic organisms—preferably plants—which comprise these systems or were generated using these methods.

An object of the present invention is to provide a transformation system for Labyrinthulomycota that allows the elucidation of biosynthetic mechanisms of lipids such as PUFA and carotenoids as well as for the construction of a high production system and the design and development of novel functional lipid molecules by the control of the mechanisms. The present invention provides a vector for the transformation of Labyrinthulomycota with a transgene, which comprises at least (1) a nucleotide sequence which is homologous to a part of chromosomal DNA of Labyrinthulomycota and is capable of homologous recombination with the chromosomal DNA, (2) a selection marker gene having a promoter sequence located upstream and a terminator sequence located downstream, and (3) a cloning site for transgene insertion having a promoter sequence located upstream and a terminator sequence located downstream.

Disclosed are DNA segments encoding <DEL-S DATE="20010821" ID="DEL-S-00001">hyaluronic acid synthase which are employed to construct recombinant cells useful in the production of hyaluronate synthase or hyaluronic acid (HA)<DEL-E ID="DEL-S-00001"> <INS-S DATE="20010821" ID="INS-S-00001">the recombinant DNA segment identified in FIG. 5. <INS-E ID="INS-S-00001">In preferred aspects, chromosomal DNA from Streptococcus equisimilis is partially digested with EcoRI and the resultant fragments are ligated to form recombinant vectors. These vectors are useful in the transformation of host cells such as E. coli and or Streptococcal hosts. <DEL-S DATE="20010821" ID="DEL-S-00002">Resultant transformants are screened by the novel screening assays to identify colonies which have incorporated HA synthase DNA in a form that is being actively transcribed into the corresponding HA synthase enzyme. These colonies may be selected and employed in the production of the enzyme itself or its product, HA.<DEL-E ID="DEL-S-00002"> <INS-S DATE="20010821" ID="INS-S-00002">The recombinant DNA segment identified in FIG. 5 is then inserted into a recombinant Streptococcal host for the production of hyaluronic acid (HA).<INS-E ID="INS-S-00002">

Methods of modifying, repairing, attenuating and inactivating a gene or other chromosomal DNA in a cell are disclosed. Also disclosed are methods of treating or prophylaxis of a genetic disease in an individual in need thereof. Further disclosed are chimeric restriction endonucleases.

embodiments of the invention include purification of DNA, preferably plasmid DNA, by use of selective precipitation, preferably by addition of compaction agents Also included is a scaleable method for the liquid-phase separation of DNA from RNA. RNA may also be recovered by fractional precipitation according to the invention. RNA, commonly the major contaminant in DNA preparations, can be left in solution while valuable purified plasmid DNA is directly precipitated. Endotoxin can also be kept to very low levels. The invention includes mini-preps, preferably of plasmid and chromosomal DNA to obtain sequenceable and restriction digestible DNA in high yields in multiple simultaneous procedures. As a method of assay, a labeled probe is precipitated by hybridizing it to a target, (erg. chromosomal DNA, oligonucleotides, Ribosomal RNA, tRNA), and thereafter precipitating the probe/target complex with compaction agents and leaving in solution any unhybridized probe.

Methods of modifying, repairing, attenuating and inactivating a gene or other chromosomal DNA in a cell are disclosed. Also disclosed are methods of treating or prophylaxis of a genetic disease in an individual in need thereof.

Methods and compositions for staining based upon nucleic acid sequence that employ nucleic acid probes are provided. Said methods produce staining patterns that can be tailored for specific cytogenetic analyses. Said probes are appropriate for in situ hybridization and stain both interphase and metaphase chromosomal material with reliable signals. The nucleic acid probes are typically of a complexity greater than 50 kb, the complexity depending upon the cytogenetic application. Methods and reagents are provided for the detection of genetic rearrangements. Probes and test kits are provided for use in detecting genetic rearrangements, particularly for use in tumor cytogenetics, in the detection of disease related loci, specifically cancer, such as chronic myelogenous leukemia (CML), retinoblastoma, ovarian and uterine cancers, and for biological dosimetry. Methods and reagents are described for cytogenetic research, for the differentiation of cytogenetically similar but genetically different diseases, and for many prognostic and diagnostic applications.

A scalable alkaline lysis process, including procedures and devices for the isolation of large quantities (grams and kilograms) of plasmid DNA from recombinant E. coli cell. Effective, contgrollable, and economical operation, and consistent low level of host chromosomal DNA in the final plasmid product. Involves a series of new unit operations and devices for cell resuspension, cell lysis, and nuetralization.

Systems and methods are disclosed for determining gene fusion by determining a fused read containing sequencing data of a portion of a fused chromosome DNA molecule; determining a predetermined point on the genome with least one mapped portion of the fused read clipped at the predetermined point (a breakpoint); identifying two mapped read portions from two breakpoints (breakpoint pair) as a potential fusion candidate; creating one or more fusion sets based on breakpoint pairs and clustering the fusion sets into one or more fusion clusters; and identifying each fusion cluster meeting a predetermined criterion as a gene fusion.

The invention relates to an alkaline pectinase genetic engineering bacteria and a construction method thereof. The construction method comprises the steps: separating chromosomal DNA from a strain for producing alkaline pectinase, designing a primer, obtaining a target gene by PCR, constructing a recombinant plasmid, expressing in Bacillus subtilis, measuring gene sequence and carrying out comparative analysis. The gene obtained and expressed in the method is different from the enzyme gene sequences which have been reported and has the differences on the composition of individual amino acids;the culture medium adopted for fermentation of the engineering bacteria is applicable to the industrial production, an expression host is protease deficient which is conducive to the separation, the purification and the stable preservation of enzyme in the late stage, and the enzyme activity of fermentation liquid of the engineering bacteria is improved by 22 times compared with the starting strain, which can achieve 330U/ml.

The present invention provides methods to protect a subject from a respiratory disorder involving an airway obstructive disease such as asthma or chronic obstructive pulmonary disease. Provided are methods to protect a subject from an airway obstructive disease using gene therapy. Methods are provided for supplying FoxA2 function to cells of the lung and airway, such as smooth muscle and epithelial cells, by FoxA2 gene therapy. The FoxA2 gene, a modified FoxA2 gene, or a part of the gene may be introduced into the cell in a vector such that the gene remains extrachromosomal or may be integrated into the subjects chromosomal DNA for expression. These methods provide for administering to a subject in need of such treatment a therapeutically effective amount of a FoxA2 gene, or pharmaceutically acceptable composition thereof, for overexpressing the FoxA2 gene. Such methods of expressing the administered FoxA2 gene in the lungs and airway provide for: (1) preventing or alleving bronchial hyperresponsiveness; (2) preventing or alleving of an airway obstructive disease, e.g., bronchial hyperreactivity, airway hyperresponsiveness, asthma or chronic obstructive pulmonary disorder (“COPD”); (3) reducing the airway resistance response to inhaled natural or synthetic bronchoconstrictors or allergens or to exercise; and (4) enhancing responsiveness (relaxation) of airway tissues to β-agonists.

The present invention is directed to methods for determining the presence or absence of a genetic defect in an IVF embryo prior to transfer comprising performing real-time PCR and 2^−ΔΔC_T analyses to determine normalized copy number of at least one invariant locus on at least one chromosome collected from at least one cell of the embryo and selecting a candidate IVF embryo determined to be without genetic defect for transfer.

The invention relates to a method of separating extra-chromosomal DNA from RNA. It also relates to DNA produced by the method and pharmaceuticals derived from such DNA, for example, DNA vaccines. The method comprises separating extra-chromosomal DNA from RNA without first digesting the RNA, and comprises: i) lysing cells comprising extra-chromosomal DNA to form a lysate; ii) clarifying the lysate; and iii) subjecting the clarified lysate to tangential flow ultra filtration under conditions, which allow a substantial amount of the RNA to pass through a membrane whilst the extra-chromosomal DNA is retained. The preferred conditions include the use of a diafiltration buffer of low conductivity and diafiltration volumes of over 10 volume equivalents.

The present disclosure relates to methods of integrating recombinant polynucleotides into genomes of unicellular organisms. In particular, the present disclosure relates to the modified unicellular organisms that contain integrated recombinant polynucleotides in their genomes and methods for production of commodity chemicals by the use of such organisms.

The present invention is expression vector to secretion express foreign gene in colibacillus or bacillus and its construction, and belongs to the field of gene engineering technology. The expression vector pBL-WZX contains one promoter sequence with function in both colibacillus and bacillus, one signal peptide sequence with function in both colibacillus and bacillus, one polyclonal site for foreign gene to be cloned, one artificially transcription termination sequence, one genetic marker for cloning selection between colibacillus and bacillus, and one DNA sequence for foreign gene to complete integration and gene copy number proliferation in the chromosome DNA of bacillus. The expression vector pBL-WZX can induce the transcription and translation of foreign gene and secrete the gene product to outside cell, and has foreign gene secretion expression interventing level up to 1 mg/mL in colibacillus and 3.1 mg/mL in bacillus separately.

Plasmid DNA is purified from a cell lysate by acidifying the lysate with a combination of a mineral acid and an inorganic salt in place of the commonly used acetate buffer, allowing chromosomal DNA and other impurities to precipitate out and recovering the supernatant, and applying the supernatant directly to a hydroxyapatite chromatographic separation medium. The use of the mineral acid and organic salt avoids the degradation of the hydroxyapatite that frequently accompanies repeated use with acetate and related compounds.

In accordance with the invention, there is provided a method for preparing plasmid from host cells which contain the plasmid, comprising the steps: (a) preparing a cleared lysate of the host cells, wherein the cleared lysate comprises unligatable open circular plasmid, wherein the open circular plasmid is not 3'-hydroxyl, 5-phosphate nicked plasmid; (b) incubating the unligatable open circular plasmid with one or more enzymes in the presence of their appropriate nucleotide cofactors, whereby the unligatable open circular plasmid is converted to 3'-hydroxyl, 5'-phosphate nicked plasmid; (c) incubating the 3'-hydroxyl, 5'-phosphate nicked plasmid with DNA ligase in the presence of DNA ligase nucleotide cofactor, whereby 3'-hydroxyl, 5'-phosphate nicked plasmid is converted to relaxed covalently closed circular plasmid; and (d) incubating the relaxed covalently closed circular plasmid with DNA gyrase in the presence of DNA gyrase nucleotide cofactor, whereby relaxed covalently closed circular plasmid is converted to negatively supercoiled plasmid. Preferably, the enzymatic steps (b), (c), and (d) are performed in a single step using an enzyme mixture comprising DNA polymerase, DNA ligase, and DNA gyrase. Preferably, the mixture further comprises a 3' terminus deblocking enzyme, such as exonuclease III or 3'-phosphatase. Preferably, the mixture further comprises one or more regenerating enzymes and a high energy phosphate donor, whereby the nucleotide by-products of the nucleotide cofactors generated by DNA ligase and DNA gyrase are converted to back to nucleotide cofactor. Preferably, the enzyme mixture further comprises one or more exonucleases, such as ATP dependent exonuclease, whereby linear chromosomal DNA is selectively degraded.