52 results about "Genetic Technique" patented technology

The present invention concerns methods and kits for diagnosing a disease condition characterized by non-physiological levels of hepcidin, comprising obtaining a tissue or fluid sample from a subject; contacting the sample with an antibody or fragment thereof that specifically binds to a polypeptide corresponding to the mid-portion or C terminus of a hepcidin protein, and quantifying the hepcidin level using an assay based on binding of the antibody and the polypeptide; wherein the non-physiological level of hepcidin is indicative of the disease condition. The present invention also concerns diagnostic methods and kits for applications in genetic technological approaches, such as for overexpressing or downregulating hepcidin. The present invention further concerns therapeutic treatment of certain diseases by treatment of subjects with hepcidin and agonists or antagonists of hepcidin.

One embodiment of the present invention provides a system that optimizes a regression model which predicts a signal as a function of a set of available signals. During operation, the system receives training data for the set of available signals from a computer system during normal fault-free operation. The system also receives an objective function which can be used to evaluate how well a regression model predicts the signal. Next, the system initializes a pool of candidate regression models which includes at least two candidate regression models, wherein each candidate regression model in the pool includes a subset of the set of available signals. The system then optimizes the regression model by iteratively: (1) selecting two regression models U and V from the pool of candidate regression models, wherein regression models U and V best predict the signal based on the training data and the objective function; (2) using a genetic technique to create an offspring regression model W from U and V by combining parts of the two regression models U and V; and (3) adding W to the pool of candidate regression models.

The present invention relates generally to recombinant genetic technology. More particularly, the present invention relates to compositions and methods for use in selection and isolation of nucleic acid molecules. The invention further relates to methods for the preparation of individual nucleic acid molecules and populations of nucleic acid molecules, as well as nucleic acid molecules produced by these methods. The invention also relates to screening and / or selection methods for identifying and / or isolating nucleic acid molecules which have one or more common features (e.g., characteristics, activities, etc) and populations of nucleic acid molecules which share one or more features.

The invention discloses a Newcastle disease virus heat-resistant transformation method and application of the Newcastle disease virus heat-resistant transformation method. The heat-resistant characteristic of the Newcastle disease virus is specifically and remarkably improved through an HN gene replacement method. The non-heat-resistant Newcastle disease virus is transformed at the transcription plasmid level through the RNA virus reverse genetic technology, the HN gene of the non-heat-resistant Newcastle disease virus is replaced with the HN gene of the heat-resistant Newcastle disease virus, host cells are transfected by the transformed transcription plasmid, rescue is conducted to obtain the heat-resistant transformed Newcastle disease virus, and therefore the heat-resistant characteristic of the Newcastle disease virus is greatly improved. The heat-resistant characteristic of the Newcastle disease virus is remarkably improved through the artificial recombination mode for the first time, and a non-heat-resistant Newcastle disease vaccine strain LaSota is successfully transformed into a heat-resistant Newcastle disease vaccine strain rT-HN. The method can be widely applied to heat-resistant transformation of Newcastle disease vaccine strains, and has great application prospects on the aspects of heat-resistant, safe and efficient Newcastle disease vaccine research and the like.

The invention provides an avian influenza virus D3 / F-R2 / 6*reassortment and construction method thereof, which is related to the field of reversal genetic technology and comprises, reassorting the six internal genes of the MPAIV chicken embryonic highly adaptive strain A / Chicken / Shanghai / F / 98(H9N2) PB2, PB1, PA, NP, M, NS and the NA gene and attenuate HA geine of the HPAIV epidemigenic strain A / Duck / Huadong / D3 / 00 (H5N1), thus constructing an avian influenza virus D3 / F-R2 / 6*reassortment, which can be used for manufacturing vaccine for treating avian influenza.

Embodiments of the present invention provides a system that optimizes a regression model which predicts a signal as a function of a set of available signals. These embodiments use a genetic technique to optimize the regression model, which involves using a portion of the sample signals used to generate each parent regression model from a pair of best-fit parent regression models to generate a child regression model. In addition, in embodiments of the present invention, the system introduces “mutations” to the set of sample signals used to create the child regression model in an attempt to create more robust child regression models during the optimization process.

The invention discloses a core primer composition for Brassica SSR (simple sequence repeats) and belongs to the technical field of molecular inheritance. The core primer composition comprises 190 core SSR primer pairs for 19 chromosomes in Brassica A and C genomes; and 10 SSR primer pairs are uniformly distributed on each chromosome, which have high polymorphism levels, substantially consistent annealing temperature and clear amplification band patterns. The core SSR primer composition is applied to increasing the molecular marking efficiency by 3 to 5 times without increasing the working costs, and is particularly suitable for the identification, the genetic diversity analysis, the nullisome and abnormal chromosome individual identification and the seed purity determination of the Brassica germplasm resources. The core SSR primer composition has the advantages of simpleness in operation, high stability and good reproducibility.

The invention discloses a method for utilizing Nsp2 gene deletion for constructing a PRRSV gene deletion vaccine virus strain and the application thereof. The method for lowering the toxicity of the PRRSV BJ-4 strain of the invention is that a nucleotide segment with the length of 69nt in a Nsp2 coding region of the PRRSV BJ-4 strain genome is continuously deleted to obtain the virus strain with reduced toxicity; the nucleotide sequence of the 69nt nucleotide segment is shown as the 2795 to 2863 sites from the 5' tail end of GenBank Accession Number AF331831. The Nsp2 of the virus rV63 which is prepared by the method for lowering the toxicity of the PRRSV BJ-4 strain deletes 21 amino acids, which has the potential to develop a marker vaccine and can identify the diagnosis method of the deleted polypeptide. When in the development process of vaccine, the invention can utilizes a reverse genetic technical platform which is already established for carrying out the transformation of GP5 glycosylation sites, thus overcoming the suppression factors which affect the neutralizing epitope and further improving the level of the vaccine-induced neutralizing antibody.

The invention provides an entire-gene cDNA clone of modified measles virus, which is obtained from the substitution and transformation of main surface antigen genes (H gene) of main existing epidemic strains (genotype of H1 and representative strain of China93-7) in China to the full-length cDNA clones of measles virus strain CC-47. The invention adopts the RNA virus reverse genetic technique to carry out the rescue of modified CC-47 virus on the cDNA level so as to obtain modified infective virus, thus providing great convenience for the research on the replication, the proliferation and the package of novel virus stains. In addition, the novel produced infective virus has the hemagglutinin antigen of an epidemic strain and has more suitable application value of the hoplivac.

The invention relates to application of a set of SNP loci in screening of populus tomentosa with characters of high photosynthesis and quick growth and a kit, and belongs to the technical field of molecular heredity. According to the application of the set of SNP loci in screening of the populus tomentosa with the characters of high photosynthesis and quick growth, the SNP loci are basic groups in the positions of 2068, 2347 and 2457 of the populus tomentosa PetC gene. Through the application of the SNP loci, quick screening of the populus tomentosa high in photosynthesis and quick in growth can be achieved, and the breeding progress is greatly accelerated.

The native chicken marking and mating technique is belongs to the field of domestic bird genetic technique. The Beijing oil chichen has golden genetic genes, phoenix head, hair feet and five toes. Take it as the paternal to cross with the high layer hen which has silver genes, and the main strain is the import high-yield mating female line hen(such as roman egg chicken, halen egg chicken and yisa egg chicken). The hens of the filial generation are having golden feathers, while the cocks are having efflorescence feathers, so the day-old chick's sex can be distinguished by the feather. The eggs produced by the hens of filial generation, basically contains the character of the crossing imitation eggs. It improves the egg-laying property substantially, reduces the cost of production, and has great economic benefit and social benefit.

The present invention concerns methods and kits for diagnosing a disease condition characterized by non-physiological levels of hepcidin protein, including prohepcidin and fragments thereof, comprising obtaining a tissue or fluid sample from a subject; contacting the sample with an antibody or fragment thereof that specifically binds to a polypeptide corresponding to the mid-portion or C terminus of a hepcidin protein, and quantifying the hepcidin level using an assay based on binding of the antibody and the polypeptide; wherein the non-physiological level of hepcidin is indicative of the disease condition. The present invention also concerns diagnostic methods and kits for applications in genetic technological approaches, such as for overexpressing or down-regulating hepcidin. The present invention further concerns therapeutic treatment of certain diseases by treatment of subjects with hepcidin and agonists or antagonists of hepcidin.

The present invention relates generally to recombinant genetic technology. More particularly, the present invention relates to compositions and methods for use in selection and isolation of nucleic acid molecules. The invention further relates to methods for the preparation of individual nucleic acid molecules and populations of nucleic acid molecules, as well as nucleic acid molecules produced by these methods. The invention also relates to screening and / or selection methods for identifying and / or isolating nucleic acid molecules which have one or more common features (e.g., characteristics, activities, etc) and populations of nucleic acid molecules which share one or more features.

The method of identifying Date Palm gender using SCAR primers includes using modern genetics techniques for detecting a novel sex-linked marker (SEQ ID NO: 1) in a Date Palm sample. The presence of the Date Palm sex-linked marker (SEQ ID NO: 1) in the sample is indicative that the sample is from a male Date Palm plant. The method of identifying Date Palm gender to determine the gender of Date Palms can be used to determine the gender of Date Palms of any age.

The invention discloses a method for genetic modification of intestinal protobacteria based on microencapsulation treatment. The method mainly includes utilizing the method of synthetic biology to genetically transform the selected intestinal primary escherichia coli and then perform microencapsulation treatment. The biosafety synthetic biology technology is combined with the non-invasive optogenetic technique for the treatment of intestinal diseases, and the method has many advantages compared with the traditional methods for treating the intestinal diseases. The intestinal primary escherichia coli are subjected to genetic modification, and the intestinal colonization compliance of the microencapsulated protobacteria is improved in the later stage. A single functional bioactive molecule,that is, a transforming growth factor (TGF-beta1) secreted by the primary escherichia coli is taken as an example to study the application in treatment of inflammatory bowel disease (IBD). The methodprovides a new idea for studying the interaction between enteric microorganisms and the body.

The invention provides a recombinant Newcastle disease virus (NDV) for expressing a VP3 gene of a new duck parvovirus and application of the recombinant NDV. The ORF of the VP3 gene of the new duck parvovirus is inserted into a full-length transcription vector pLMV-RFP of the NDV by utilizing a reverse genetic manipulation platform of an established NDV PHY.LMV42 strain, so as to construct a recombinant plasmid pLMV-NDPV-VP3, and the recombinant plasmid and three auxiliary plasmids are together transfected with BHK-21 cells to obtain a recombinant vaccine strain rLMV-NDPV-VP3 which is higher in reproductive performance and capable of expressing NDPV-VP3 protein. The NDPV-VP3 gene is located in a non-coding region between a P gene and an M gene of the NDV, and a recombinant virus rLMV-NDPV-VP3 is obtained through reverse genetic technique. The recombinant NDV for expressing the VP3 gene of the new duck parvovirus can be used for preventing duck new parvovirus disease and duck Newcastledisease.

The invention relates to a cloning method of an influenza virus gene, in particular to PCR (polymerase chain reaction) amplification primers of an influenza A virus gene and a rapid cloning method of the influenza A virus gene. According to the PCR amplification primers of the influenza A virus gene and the rapid cloning method of the influenza A virus gene, the PCR amplification primers of eight gene segments of the influenza A virus and primers of linearized influenza virus reverse genetic vectors are provided and the rapid cloning method of the influenza A virus gene is further provided, the influenza virus gene segments and the linearized influenza virus reverse genetic vectors are amplified by the primers through PCR, and then the influenza virus gene segments and the linearized influenza virus reverse genetic vectors are recombined and cloned in vitro by recombinase and transformed to escherichia coli for cloning. According to the invention, the gene cloning process of a traditional influenza virus reverse genetic technology is greatly simplified, and the problems of complex influenza virus gene cloning process and low efficiency of the traditional influenza virus reverse genetic technology are solved.

The invention provides a vaccine strain rSHA-[delta]200, and a construction method and application thereof, relates to that field of influenza vaccine. The vaccine strain rSHA-[delta]200 includes a HAgene that does not contain an NRT glycosylation site. The invention finds out the potential glycosylation sites on the HA protein through analysis, deletes glycosylation sites by site-directed mutagenesis technology, constructs HA expression plasmids with multiple glycosylation sites deleted, obtains recombinant vaccine strains by using 8 plasmid reverse genetic technology, and prepares antiserum. The rSHA-[delta]200 obtained in the invention is an ideal candidate for inactivated vaccine against H9 subtype AIV because of its good cross-immunity and serum cross-neutralization ability.

The invention aims to provide a recombinant canine measles virus expressing luciferase NLuc. The structure of a recombinant CMV nucleic acid fragment for rescuing and expressing NLuc is 5'-N-P-NLuc-M-F-H-L-3', wherein N, P, M, F, H and L are structural genes of the virus; NLuc is ORF of luciferase, the upstream of the ORF contains an M gene initiation sequence and a Not I restriction enzyme cutting site, and the downstream contains a Pme I restriction enzyme cutting site and a P gene termination sequence. The rCMV-NLuc rescued by a reverse genetic technology has a high expression level on NLucin cells, and can emit macroscopic light blue fluorescence under the action of a luminescent substrate furimazine. The insertion of an exogenous gene does not significantly affect the growth kineticsof the virus, and the exogenous gene is relatively stable in the virus passage process.

The method to modify wild NPV and construct the new for Lepidoptera pest with anti-sense- genetic technique is presented and comprises the scheme as: (1) cloning CS conservative sequence of geometrid; (2) constructing the transfer carrier for the anti-sense CS gene of Lepidoptera pest; (3) recombining NPV; (4) validating the NPV with PCR or other methods; (5) contrast with wild NPV, evaluating the insect disinfectation capacity of recombined NPV for screening the virus strain with higher virulence and faster diffusion and secondary infection speed.

The invention provides a mink enteritis parvovirus whole genome infectious clone and a construction method and application thereof. The method comprises the following steps of: carrying out enzyme digestion on a complete genome in an MEV replication form, and sequentially and directionally cloning to a vector pUC18 M in a segmented form to obtain a recombinant plasmid; and mixing the recombinantplasmid with a transfection reagent, and transfecting CRFK cells to obtain a rescue virus. The mink enteritis parvovirus infectious clone constructed by utilizing a reverse genetic technology transfects the CRFK cells in vitro, and can induce the cells to generate cytopathic effects and growth trends the same as those of a parent virus. The method is simple, rapid, time-saving and labor-saving, and has higher accuracy compared with a traditional PCR method, and base mutation can be rapidly and conveniently carried out at any position of the MEV genome by applying the cloning system, so that aneffective way and means are provided for subsequent research and development of molecular biology of MEV and research and development of vaccines.

The invention provides a method for isolating or enriching a rare cell from a biological fluid of a mammal employing an antibody that binds a cell-surface antigen of the rare cell. The immobilized antibody is incubated with a sample of biological fluid that includes the rare cells and a plurality of other cells so as to form an antibody-rare cell complex. The complex can be detected or isolated and subsequently analyzed by any of a variety of physical, chemical and genetic techniques.

In the present invention, a recombination of gene groups of nemadectin aglycon biosynthesis is performed for obtaining C-13 hydroxylnemadectin, to which sugar groups can be attached, and a production strain which produces C-13 hydroxylnemadectin is produced. Further, C-13 glycosylnemadectin producing strain is prepared by introducing aveBI-BVIII genes involving glycosidation of avermectin and biosynthesis of oleandrose. As described, C-13 hydroxylnemadectin and C-13 glycosidated nemadectin can be obtained effectively by using the producing strain prepared by means of the molecular genetic technology, and improvement in the biological activity thereof can be expected.

The method of identifying Date Palm gender using SCAR primers includes using modern genetics techniques for detecting a novel sex-linked marker (SEQ ID NO: 1) in a Date Palm sample. The presence of the Date Palm sex-linked marker (SEQ ID NO: 1) in the sample is indicative that the sample is from a male Date Palm plant. The method of identifying Date Palm gender to determine the gender of Date Palms can be used to determine the gender of Date Palms of any age.

The present invention relates to a mutated virus. Said virus can be an influenza virus of human or other animal origin. The present invention also relates to a method for preparing the mutated virus, the method comprising introducing UAG codons into positions upstream of the stop codons per se of one or more genes of a viral genome by reverse genetic techniques. The present invention further relates to uses of the mutated virus, for example, as a live attenuated vaccine, or in replication of controllable and safe virus models, and the like.

The invention discloses an oncolytic virus NDV-NRP1 and a construction method and application thereof, and belongs to the technical field of microorganisms. A recombinant virus NDV-NRP1-scFv-GT is constructed by adopting a reverse genetic technology, specific replication of rNDV in tumor cells is achieved at the transcriptional level by using an hTERT promoter, by combining NRP1-scFv with NRP1, invasion and growth of tumors are inhibited, and alpha-Gal epitope is overexpressed, so that anti-Gal of a human body can be induced to specifically target the tumor cells, complement is activated, killing of the tumor cells is achieved, and an effective anti-tumor effect is achieved.

The invention discloses a method for quickly constructing an IBV (Avian Infectious Bronchitis Virus) reverse genetic strain, and belongs to the technical field of coronavirus reverse genetics. The constructing method comprises the following steps: quickly completing construction containing IBV genomic full-strength cDNA (Complementary Deoxyribose Nucleic Acid) clone by taking a BAC (Bacterial Artificial Chromosome) vector as a framework and applying an in-vitro homologous recombination technology, directly transfecting cells by a constructed recombinant plasmid, and transcribing in the cells, thus obtaining a transcript having infectivity; completing virus packaging; inoculating SPF (Specific Pathogen Free) chick embryo to a mixed solution of the cells and a culture medium and passing from generation to generation, thus obtaining the IBV reverse genetic strain. The constructing method disclosed by the invention has the advantages of simple operation and high positive cloning efficiency, the obtained IBV reverse genetic strain has passage stability, and an effective tool is provided for researching pathogenesis of the virus in vitro, developing a novel vaccine and the like; according to the method disclosed by the invention, transcription is carried out in the cells by utilizing a CMV (Cytomegalovirus) promoter added on a 5' terminal, and the rescue efficiency of the virus is greatly increased by utilizing an HDVR (Hepatitis Delta Virus Ribozyme) sequence added on a 3' terminal.