292 results about "Function genes" patented technology

RNAi arrays and methods for using the same are provided. The subject arrays are characterized by having two or more distinct RNAi agents. The arrays find use in methods where cells are contacted with the arrays and the activity of the RNAi agents is determined by evaluating the contacted cells. The subject arrays and methods find use in a variety of applications, such as high throughput loss of function genomic applications.

The invention discloses a method for improving the efficiency of CRISPR mediated homologous recombination. The method includes the steps of: inhibiting the expression of Lig4, DNA-PK and XRCC6 by shRNA so as to inhibit the non-homologous end joining (NHEJ) repair pathway; conducting fusion expression on sgRNA and shRNA at targeting genome specific positions to form shRNA-sgRNA polycistron; and placing the polycistron at the RNA polymerase II or RNA polymerase III promoter downstream. The carrier constructed by the method provided by the invention has a concise structure and is convenient to use, also enhances the homologous recombination efficiency by dozens of times, greatly expands the function and application of the CRISPR carrier, and is a good tool for functional gene and signal pathway research, medical research, and recombinant protein expression.

Isolated polynucleotides, and vectors including the same, are disclosed as useful for down-regulation of specific RNA in cells, including a first sequence of about 17 to about 23 nucleotides, complementary to said RNA, and linked to a second sequence capable of forming a loop when said second sequence is RNA. The polynucleotides include self-complementing single-stranded polynucleotides, including a third sequence linked by said second sequence where all nucleotides in said first and said third sequences are complementary. Functional genomic, diagnostic and therapeutic methods are disclosed that involve reducing the amount of a unique RNA sequence in cells using a vector encoding the self-complementing polynucleotide including a first sequence complementary to said RNA sequence. Methods are also disclosed for preparing the polynucleotides, vectors, libraries of vectors, and the temporary knock-down of proteins, such as lethal proteins, during virus or recombinant protein production.

Methods are provided for the rapid identification of essential or conditionally essential DNA segments in any species of haploid cell (one copy chromosome per cell) that is capable of being transformed by artificial means and is capable of undergoing DNA recombination. This system offers an enhanced means of identifying essential function genes in diploid pathogens, such as gram-negative and gram-positive bacteria.

The invention provides a group of oligonucleotide aptamers Apt-1, Apt-2 and Apt-3 which are capable of simultaneously identifying clenbuterol hydrochloride and salbutamol, an oligonucleotide aptamer CLB-2 which is capable of identifying the clenbuterol hydrochloride with high specificity, an oligonucleotide aptamer SAL-5 which is capable of identifying the salbutamol with high specificity and two oligonucleotide aptamers RAC-5 and RAC-6 which are capable of identifying ractopamine with high specificity. Through an SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technology based on Fe3O4 magnetic nanoparticle separation, a random oligonucleotide library is immobilized on avidin-enveloped magnetic nanoparticles by virtue of a complementary chain of a biotinylation marker, and the oligonucleotide aptamers, which are high in specific affinity, are finally obtained by conducting screening by 16 turns. The aptamers are broad in application prospect; and the aptamers, by virtue of marker function genes or fluorescent dyes, are applicable to detection of the clenbuterol hydrochloride, the salbutamol and the ractopamine in food; therefore, a new choice is provided for existing detection methods which depend on antibodies.

The invention belongs to the field of establishment of a population genetic resource bank, and provides a scientific establishment method of a goat resource population bank, and provides a goat reference family system establishment case, and relates to two national-level genetic resource varieties, namely, a Dazu black goat and an Inner Mongolia Cashmere goat (Alxa type). According to the resource population establishment method, forward cross and reciprocal cross of Dazu black goat and Inner Mongolia Cashmere goat (Alxa type) are utilized, random mating of genetic relationship is avoided for F1, so that Mendel genetic characters are separated, a goat economic character QTL or a functional gene is located by virtue of a genome means, and a relatively-precise genetic mark is provided for the goat breeding. The genetic resource bank comprises goat population, a genealogy inforamtion database, a phenotypic character database, a DNA sample library, an RNA sample library and a genetic variation information database.

The invention relates to a construction and an application of recombinant Chicken Marek's Disease Virus SC9-1 strain and SC9-2 strain. The recombinant virus construction method solves the technical problem that kanr gene cannot be knocked out again by a present method after kanr gene containing flp recognition sites at two ends is continuously used twice to knock out two specific functional genes on the same virus genome. The obtained recombinant virus MDV SC 9-1 strain and MDV SC 9-2 strain are used as production strains of Marek's Disease live vaccine. The prepared vaccine prevents the very virulent or emerging very virulent plus MDV induced chicken Marek's disease. The protective immunity effect of the vaccine is superior to CVI988/Rispens strain vaccine which is mostly widely used in foreign and domestic markets at present. The antigenicity of the recombinant virus is more similar to that of Chinese epidemic strain than the antigenicity of the similar virus rMd5deltameq which has been published in the United State. The strains provided by the invention will not induce tumor and has no immune suppression effect. Therefore, the strains are more applicable to China.

The invention discloses a novel capture and enrichment technology for targeting nucleic acid molecules. The targeting nucleic acid molecule fragments are hybridized through special capture probes; and probe molecules marked with acrylamide groups are fixed in acrylamide glue, so that the targeting nucleic acid molecule fragments are captured. The technology can be used for functional gene capture and high-throughput sequencing analysis of microbial community, fishing and high-throughput sequencing analysis of genes related to diseases in human genome, and fishing and analysis of signature sequences of environment pathogenic microorganisms. Besides, the technology can be used for development of diagnostic kit for a plurality of molecular diseases and pathogenic microorganisms. The technology is relatively low in cost. Glue efficiency of acrylamide molecules is far higher than connection efficiency of magnetic beads and probes, so that capture efficiency is greatly increased.

The invention discloses a multifunctional supramolecular gene delivery system based on polyethyleneimine-cyclodextrin and a preparation method thereof. A polyethyleneimine-cyclodextrin polycationic compound serves as a framework, adamantane-disulfide bond-polyethylene glycol having oxidation reduction reactivity and adamantane-polyethylene glycol-targeting peptide having targeting performance serve as modified groups, polyethyleneimine-cyclodextrin is combined with adamantane-disulfide bond-polyethylene glycol and adamantane-polyethylene glycol-targeting peptide through a self-assembling principle, and a subjective body and an objective body act to form a supramolecular gene carrier. The delivery system is multifunctional, has oxidation reduction sensitivity, specificity for targeting liver cancer and excellent gene transfection efficiency, and can carry functional genes to efficiently treat liver malignant tumor.

The invention discloses a method and application for promoting cells to secrete exosomes, and relates to the fields of biological materials, tissue engineering and regenerative medicines. According tothe method, the cells are stimulated by active signals of a biological material, and the exosomes generated by the cells are collected, and are used for tissue wound treatment. The operation route ofthe method and the application scheme of the obtained exosomes are described in detail, under the stimulation effects of bioactive glass chemical signals, electrostatic spinning fiber membrane structure signals and chemical/structural combination signals, more exosome particles can be secreted by mesenchymal stem cells and umbilical vein endothelial cells, and the exosomes can promote the corresponding target cells to express higher levels of functional genes, secrete more multifunctional proteins and carry out deep functional differentiation, so that the tissue wound surface can be rapidly repaired. The exosomes obtained by the method disclosed by the invention have the characteristics of higher yield and stronger tissue repair promotion function.

The invention relates to a method for converting disease-resistance genes of rice and obtaining transgenic descendants without selective markers. The method comprises the following steps: first, cloning green fluorescent protein (GFP) and hygromycin phosphotransferase (HPT) on a marker gene carrier, placing a target gene in other T-DNA carrier, mixing strains of agrobacterium tumefaciens carrying the marker gene carrier and strains of agrobacterium tumefaciens carrying the target gene carrier and converting callus of the rice, performing PCR (polymerase chain reaction) detection on the disease-resistance genes of the target gene carrier by a specific primer, and screening to obtain co-transformation plants (T0); then, screening the marker gene plants with positive GFP rapidly and massively by means of a desk lamp fluorescence detector in segregative generations (T1 or T2) of the co-transformation plant for removing, and performing PCR detection for disease-resistance genes to the plants with negative GFP so as to obtain the individuals without selective markers of the transgenic disease-resistance genes. The method can be applied to transgenic breeding without selective markers of rice blast-resistant genes or other functional genes of the rice, and enhances the disease resistance of the rice or improves other agronomic traits.

The invention discloses flavoprotein oxidase genes of macleaya cordata in synthesis of sanguinarine and chelerythrine and application of the flavoprotein oxidase genes. Three flavoprotein oxidase genes joining in synthesis of sanguinarine and chelerythrine are found in a macleaya cordata genome for the first time, including genes Mc20113, Mc6407 and Mc6408. Steps of reaction are respectively verified by using a brewer's yeast system through upstream precursor feeding, and synthesis of sanguinarine and chelerythrine and midbodies can be achieved. The conversion efficiency of same functional genes of macleaya cordata and opium poppy is further compared by using a brewer's yeast heterologous expression system, and the result shows that the conversion rate of the gene of macleaya cordata is remarkably higher than that of opium poppy. The invention further discloses a molecular mechanism of synthesis of sanguinarine in macleaya cordata, and a theoretic basis and molecular assistant breeding targets are provided for breeding of macleaya cordata with high contents of sanguinarine and chelerythrine; and meanwhile, precious experience is provided for in-vitro artificial synthesis of sanguinarine and chelerythrine.

The present invention relates to a high sensitivity, high fidelity, high uniformity and high efficiency amplification technology of trace and complex DNA. The technology system is constant temperature DNA amplification optimized by high concentration trehalose and guided by DNA polymerase, wherein balanced and efficient amplification of DNA in different regions of the whole genome or a DNA library sequence can be maintained in the context of effective inhibition on non-specific amplification by the amplification system. The method has the following characteristics that: sensitivity is high; balanced amplification of different region sequences can be maintained, and the amplification product has high fidelity (minimum deviation) and high uniformity; and the total reaction volume has high flexibility, and can be conveniently adjusted according to requirements. In addition, the technology provides broad application prospects in single cell level and trace genome DNA amplification and gene detection, pretreated function genome associated DNA library (spectrum) analysis, and other fields.

A method for detecting transcription factor protein by nucleic acid probe with nuclease protection PRET label includes preparing fluorescence labeled nucleic acid probe, carrying out combination reaction of nucleic acid probe with transcription factor protein, adding nuclease in combination reaction system to carry out enzyme nick reaction and detecting reaction system by fluorescence to reflect expression and activation level of transcription factor protein.

A binary expression carrier of Agrobacterium for culturing the seeds of plant by genetic engineering contains the closely linked corm ubiquitin promoter Pubi regulated Arabidopsisí» cold response gene transcription activation factor gene cbf1 and the Arabidopsis specific senescence gene promote SAG12 regulated iso-pentenyl transferase ipt geneí»s two-valence stress-resistant gene. Its preparing process includes such steps as inserting the T-Nos of sag12-ipt fusion gene into a binary carrier, inserting ubi-cbf1 fusion gene and inserting sag12-ipt fusion gene.

The invention belongs to the field of molecular biology DNA labeling technology and application, and in particular relates to Apostichopus japonicas express sequence tag EST micro-satellite label screening development and application. The invention provides Apostichopus japonicas AjE101 micro-satellite specific DNA primers, a polymerase chain reaction (PCR) reaction system by utilizing the primers, and a detection method for an Apostichopus japonicas AjE101 micro-satellite DNA label. The detection method comprises the following steps of: extracting genome of Litopenaeus vannamei Boone; designing the specific primers at two ends of an Apostichopus japonicas AjE101 micro-satellite DNA core sequence; and performing PCR amplification on genome DNA of different groups of the Litopenaeus vannamei Boone or individuals in the group by using the primers, analyzing products, and determining the genome of each individual so as to obtain an Apostichopus japonicas polymorphism genetic variation map. The invention is mainly applied to Apostichopus japonicas germ plasm resource and genetic diversity analysis, cular population genetics, construction of genetic maps and research of functional genes.

The invention relates to the biochips field, in particular to a tissue chip used for functional genome research and the preparation method as well as the application thereof. The tissue chip of the invention includes a substrate and a paraffin section arranged on the surface of the substrate, wherein, the paraffin section is provided with tissue sample points which are in different development stages and array according to the tissue dynamic development process. Because the tissue chip of the invention can be combined with a gene chip technology, the space-time expression of a difference expressive gene screened out by the chip technology in the tissue dynamic development process is validated and researched in the high-quality, economic and available way.

The invention discloses a major QTL and SNP molecular marker for controlling Nelumbo nucifera color traits as well as a detection primer and application thereof. The invention firstly provides a majorQTL site for controlling the flower color traits of Nelumbo nucifera, and the major QTL site is positioned in a sixth linkage group and between two SNP markers; the major QTL is closely linked with the SNP molecular marker, has a relatively high contribution rate to the flower color traits of the Nelumbo nucifera, participates in regulating and controlling the phenotype of the red or white flowerof the Nelumbo nucifera, and can be used for map-based cloning mining of functional genes for controlling the flower color traits and molecular marker-assisted selection. The invention further provides an SNP molecular marker closely linked with the major QTL, and the SNP molecular marker is used for efficiently selecting Nelumbo nucifera varieties with target flower colors and molecular marker assisted breeding of Nelumbo nucifera with seeds and Nelumbo nucifera with roots. The invention further provides a PARMS primer group for detecting the SNP marker and a method for identifying the red and white flower traits of the Nelumbo nucifera, so that the flower color genotype data of the Nelumbo nucifera can be analyzed and obtained only through PCR amplification and without electrophoresis detection.

The invention discloses a pear hexose transport protein gene PbHT1 and application thereof. The hexose transport protein gene PbHT1 is separated from a pear fruit, the nucleotide sequence of the gene is as indicated in SEQ ID No. 1, and the coded amino acid sequence of the gene is as indicated in SEQ ID No. 2. The gene PbHT1 is transformed into a tomato through a southern buddhism genetic transformation method, a transgenic plant is obtained, and through biological function verification, it is shown that the clonal gene PbHT1 has the effects of postponing the flowering phase and raising the hexose content in the fruit. Through finding of the gene PbHT1, a new gene resource is provided for molecular breeding for improving quality of the plant fruit.

The invention discloses a method for developing polymorphic SSR markers based on transcriptome sequences. The method comprises the following steps: S1, the transcriptome sequences of a plurality of samples of a target species are obtained; S2, potential SSR site information is detected; S3, the types of SSR locus repeats and adjacent nucleotide sequence information of the SSR locus repeats are obtained by screening; S4, polymorphic SSR candidate loci among the samples are screened; and S5, transcriptome sequence sets carrying the polymorphic SSR loci are spliced into a non-repetitive contig Uni-contig, primers are designed according to a side wing conservative sequence of the non-repetitive contig Uni-contig, and marker polymorphic detection is conducted. The polymorphic SSR markers can beeffectively screened through the transcriptome sequences, the configuration requirement for a computer is reduced, the method can be completed by a common computer, the effectiveness of the molecularmarker primers is effectively improved, the development time is shortened, the development cost is reduced, meanwhile, the PCR amplification fragment length of the polymorphic SSR markers and linkagegene function information of the polymorphic SSR markers can further be predicted, the development efficiency of the transcriptome SSR markers is remarkably improved, and molecular assisted breedingand functional gene research are promoted.

The invention discloses a bacillus gene traceless knockout/knockin plasmid and method, and a kit. The genome of the bacillus gene traceless knockout/knockin plasmid contains only one kind of resistance gene marked with a positive selection marker, a replicon capable of being duplicated in escherichia coli, a thermo-sensitive type replicon capable of being duplicated in bacillus, at least one I-Scel enzyme cutting site and only one kind of chromogenic protein gene, wherein promoters in front of the resistance gene and the chromogenic protein gene are both constitutive promoters in bacillus. The invention further provides a helper plasmid used for improving the knockout/knockin efficiency of the bacillus gene traceless knockout/knockin plasmid, and resistance gene and chromogenic protein gene in the genome of the helper plasmid are different from those of the knockout/knockin plasmid. By the adoption of the two plasmids, one or more target DNA sequences in the genome of bacillus can be modified continuously or iteratively, and the plasmids can be applied to multiple fields including bacillus genetic modification, metabolic process researching, functional genome researching and industrial application researching.

The invention discloses a method for the extraction and the purification of microbial total DNA of a compost and a buffer solution thereof, the method comprises the following steps of: adding a dehumification solution in an amount of 8-15ml/g of the compost into a sample of the compost in which the DNA is to be extracted, for dehumification; then adding a DNA extraction buffer solution in an amount of 1.0-1.5ml/g of the compost for the crude extraction of the microbial total DNA; and finally, purifying the crudely-extracted DNA solution with a kit so as to obtain the microbial total DNA of the compost. The inventive method can reduce the influence of humic acids and effectively improve the yield and the purity of the DNA extracted from the compost, thus better facilitating compost microbial molecular ecology and researches of other functional genes.