728 results about "Functional genes" patented technology

Methods for preparing a biochip are provided herein wherein the biomolecular probe to be used with the biochip is alternatively bound to a hydrogel prepolymer prior to or simultaneously with polymerization of the prepolymer. In particularly preferred embodiments, a polyurethane-based hydrogel prepolymer is derivatized with an organic solvent soluble biomolecule, such as a peptide nucleic acid probe in aprotic, organic solvent. Following derivatization of the prepolymer, an aqueous solution, for example sodium bicarbonate, preferably buffered to a pH of about 7.2 to about 9.5, is added to the derivatized prepolymer solution to initiate polymerization of the hydrogel. Alternatively, a water soluble biomolecule, such as DNA or other oligonucleotide, is prepared in an aqueous solution and added to the polyurethane-based hydrogel prepolymer such that derivatization and polymerization occur, essentially, simultaneously. While the hydrogel is polymerizing, it is microspotted onto a solid substrate, preferably a silanated glass substrate, to which the hydrogel microdroplet becomes covalently bound. Most preferably the hydrogel microdroplets are at least about 30 mum thick, for example about 50 mum to about 100 mum thick. The resulting biochips are particularly useful for gene discovery, gene characterization, functional gene analysis and related studies.

The present invention relates to diagnostic methods for assessing predisposition of a subject to a mental disorder phenotype having an association with an at-risk allele of a brain-functional gene having a plurality of alleles, the association being conditioned by a pathogenic environmental risk factor status condition. Additionally, the invention relates to methods for discovering a conditional association between a mental disorder phenotype and an at-risk allele of a brain-functional gene having a plurality of alleles, the association being conditioned by a pathogenic environmental risk factor status condition.

The in vivo siRNA expression system according to this invention is a system that intracellularly expresses small interfering (si) RNAs and comprises antisense and sense code DNAs coding for antisense and sense RNAs targeting any region of a target gene mRNA and one or more promoters that function to express the antisense and sense RNAs from the antisense and sense code DNAS, respectively.

A gene activation / inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent β-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebrate development. The FLP recombination system of the present invention can be incorporated in transgenic, non-human mammals to achieve site-specific integration of transgenes, to construct functional genes or to disrupt existing genes.

A method of packaging a recombinant viral vector is carried out by: (a) providing a packaging cell, the packaging cell containing and expressing a nucleic acid encoding a mutant adenovirus E4orf6 protein, the E4orf6 protein containing at least one mutation that renders the protein non-toxic to the host cell; (b) transfecting or infecting the packaging cell with a nucleic acid that encodes a recombinant viral vector (e.g., an adenovirus vector or an adeno-associated virus vector), where the vector lacks a functional gene encoding E4orf6 protein; (c) culturing the transfected cells; and then (d) collecting packaged recombinant viral vector from the cultured cells. Nucleic acids, vectors and packaging cells used for carrying out the methods, as well as proteins utilized in the methods, are also described.

The invention provides a method for single cell classification and screening and a device therefor. The method comprises the following steps of carrying out comparison of reads obtained by sample sequencing and a reference genome, carrying out data filtration of a comparison result, determining a consistent genotype of all single cell samples according to filtered data, saving the consistent genotype of the all single cell samples into a SNP data set, extracting genotype files of loci corresponding to reference genome SNP data set positions from the saved SNP data set, selecting a cell mutation SNP locus, and carrying out cell classification and functional gene screening according to a genotype file of the cell mutation SNP locus. The method and the device avoid cell marking, solve the problem that the traditional single cell classification method can not realize classification of a certain cell subset having no corresponding specific markers, realize complete analysis of genetic variation information of a single cell genome, and greatly improve the accuracy of cell subset classification.

The invention discloses an efficient specific sgRNA recognition site guide sequence for pig gene editing and a screening method thereof. The screening method includes: screening functional genes, performing ORF analysis, predicting a functional gene sgRNA recognition site guide sequence, detecting whole-genome off-target sites, grading predicted target sites according to off-target information and target site positions, sequencing, screening and statistically counting results, optimizing algorithms and developing software. The efficient specific sgRNA recognition site guide sequence and the screening method thereof have the advantages that the pig specific sgRNA recognition site guide sequence is obtained through strict screening and inspection and includes the sgRNA recognition site guide sequences, for CRISPR-Cas9 gene editing, of all pig protein encoding genes; the authenticating, grading and inspecting algorithms for specific sgRNA recognition and software corresponding to the algorithms and used for predicting and evaluating the pig functional gene sgRNA recognition site guide sequence are widely applicable to the sgRNA specific site prediction of non-model species with whole-genome sequences.

An immune bead is made up of the magnetic carrier micro ball which combines at least one immune matching base. The micro ball is composed of the magnetic nm particle and the high molecular framework material which the core is the metal particle; the out of the core is high molecular framework, the out layer is the functional layer which can combines functional gene of different immune matching base. The manufacture method includes: the bead pretreatment, the bead activation, manufacture of the coupling antibody, closing the antibody with the confining liquid and purifying the immune bead. The detecting method is to detect the different things by the immunological response sandwich, the competition and the indirect method and set the control system on the testing board. The board is made up of the encrusting test paper, the coupling mat, the sample mat, the water suction mat, the coving film and the testing board outside calipers. It has the high sensitivity and accurate quantity; the regent is simple and cost low.

The invention relates to the technical field of genetic engineering, and discloses a recombinant yeast strain as well as a construction method and an application thereof. The recombinant yeast strain has the knockout gene gal1, gal7, gal10 or gal80, and the knockout gene ypl062w, and contains 2-4 gene segments integrated to the genome through the yeast homologous recombination. According to the invention, the yeast strain with the knockout genes is established, an optimized host cell is provided for producing lycopene, and functional genes crtE, crtB and crt I with different sources for synthetizing lycopene, and the specific yeast endogenous genes are selected, and integrated to the genome of the yeast strain with the knockout gene through the modular design, and thus a brand-new recombinant strain with high yield of lycopene is obtained.

The invention discloses a primer set for detecting functional genes of wheat on the basis of a KASP [competitive allele specific PCR (polymerase chain reaction)] technology and application of the set primer. The primer set comprises totally 14 groups of KASP primers. The KASP primers are respectively designed for the 14 functional genes of the wheat, and particular nucleotide sequences of the KASP primers are sequences 1-42 in sequence tables. The primer set and the application have the advantages that the functional genes of different wheat varieties can be quickly detected by the aid of the KASP primer set, methods for detecting the functional genes of the wheat are simple, convenient and speedy, and detection results are accurate and reliable; the primer set has an important theoretical significance and important economic value in assistant selection of the wheat varieties by the aid of molecular markers.

The invention discloses bacillus amyloliquefaciens JK6 and biological fertilizer and application. The serial number of strain preservation is CGMCC No. 10658 and is stored in the China general microbiological culture collection center on March 23, 2015. The bacillus amyloliquefaciens JK6 is mainly used for biological prevention and control of pseudomonas solanacearum and is provided with an iron-production carrier, a biological membrane, protease, catalase, cellulose and IAA characteristics, multiple fungal pathogens of banana wilt pathogen, litchi anthrax germs, litchi frost phytophthora pathogens, rice blast fungus, cucumber wilt pathogen and the like, and growth of lettuce plants is obviously promoted; the K6 strain can be successfully planted, and disease preventing functional genes of yndj, srfAB, fend and ituC are expressed in soil; the strain is high in reproduction speed, simple in production technology, high in adverse resistance, easy to store and favorable for industrialized production, the biological fertilizer is prepared with the strain, soil-born diseases can be prevented and controlled, growth of the plants can be promoted, and application prospect is broad.

The invention relates to the technical field of gene engineering, and discloses a recombinant yeast strain, and a construction method and application thereof. In the recombinant yeast strain, gal1, gal7, gal10 and ypl062w genes are knocked out, and the recombinant yeast strain comprises 9 gene segments which are integrated to the genome by yeast homologous recombination. The construction method comprises the following steps: constructing a four-knock-out yeast strain to provide an optimized host cell for producing beta-carotin, selecting lycopene cyclases crtY from different sources, and carrying out modular design to integrate functional genes crtE crtB and crtI of the specific-source synthetic lycopene, specific yeast endogenous genes and the like to the four-knock-out yeast strain genome, thereby obtaining a brand-new recombinant strain capable of producing beta-carotin at high yield.

The invention provides a functional molecular marker for rice anti-blast gene Pi9 and application thereof, belonging to the field of crop molecular genetic breeding science. The invention finds out that the rice anti-blast gene Pi9 has a 10bp insertion / deletion site positioned between the gene promoter -516 and -517, wherein the nucleotide sequence is disclosed as SEQ ID NO.1; and on such basis, the invention develops a method of the functional molecular marker for gene Pi9. By detecting the functional molecular marker, the invention can accurately detect whether genomes of different rice species contain the Pi9 functional gene and detect the homozygotic state, and can be used for screening rice hybrid transformed descendant plants to enhance the breeding efficiency of the rice anti-blast material, thereby effectively controlling the size of the breeding population, obviously saving the breeding and screening cost and obtaining the anti-blast rice species containing the Pi9 functional gene.

The invention relates to the technical field of gene engineering, and discloses a recombinant yeast strain, and a construction method and an application thereof. The recombinant yeast strain is obtained through the following steps: knocking out gal1, gal7, gal10 and ypl062w genes, integrating three gene fragments a genome through yeast homologous recombination, further knocking out an rox1 gene, and further integrating two gene fragments to the yeast genome. The gene knockout yeast strain constructed in the invention provides optimized host cells for production of lycopene; and combined functional genes crtE, crtB and crtI having specific sources and used for synthesizing the lycopene and specific yeast endogenous genes are selected and are integrated to the gene knockout strain genome through modular designing, so the brand new recombinant strain for highly yielding lycopene is obtained.

The invention provides a method for constructing a high-flux simplified genome sequencing library. The method comprises the following steps: digesting genome DNA of different samples by using restriction enzymes, thereby obtaining blunt-end DNA fragments with phosphorylation modification at the 5' end; adding the base A at the 3' end, connecting the DNA fragments of different samples with 5' phosphorylation and 3' cohesive ends A with joints containing different bar code sequences, connecting the products PCR, purifying and mixing the amplification products, performing low-cycle PCR amplification, thereby obtaining a target fragment amplification product of which the ratio of the joints to joint dimers is greatly reduced; and separating, purifying, and forming the high-flux simplified genome sequencing library from the purified products. The method disclosed by the invention has the advantages that the flux is high, the cost is low, and the generated library is high in sequencing quality and has very wide application prospects in researches such as high-flux SNP detection and marker development, genetic map drawing and functional gene location.

The invention relates to a method for obtaining gene information and functional genes from species without genome reference sequences. The invention also discloses a method for obtaining the gene expression profile of the Asian corn borer. The invention also discloses a method for obtaining transcriptome information and functional genes of the Asian corn borer. The present invention combines DGE-tag technology with RNA-seq technology for the first time to obtain gene expression period, expression level, corresponding metabolic pathway and gene function information from species without genome reference sequence. The method is convenient and fast , accurate and low cost.

The invention belongs to the field of crop biotechnology and in particular relates to a molecular marker of a rice-blast-resistant gene Pi2 and application of the molecular marker. The upstream and downstream molecular markers are based on specific insertion / deletion loci polymorphism of nucleotide sequences shown in SEQ ID NO.1 and SEQ ID NO.2. The two molecular markers are co-dominant molecular markers developed for close linkage regions on two sides of the functional gene Pi2, and have the advantages of being high in accuracy and capable of being widely applied to different breeding parents. By detecting the two molecular markers, whether a detected rice material has the rice-blast-resistant gene Pi2 or not can be accurately judged. By virtue of a molecular marker combination, whether a hybrid transferred progeny plant genome of a parent of the functional gene Pi2 and other parents has the functional gene Pi2 or not and the homozygous state of the functional gene Pi2 can be accurately detected, and the breeding efficiency of a rice-blast-resistant rice variety with the functional gene Pi2 is improved.

A method for producing a mutant gene encoding a functional gene product comprising: (a) introducing one or more mutations into a gene to form a mutated gene; (b) providing the mutated gene to a host microorganism; (c) culturing the host microorganism containing the mutated gene under conditions which allow expression of the mutant gene product; and (d) selecting a host microorganism capable of producing a functional gene production from the mutated gene.

The invention discloses a method for preparing a salvia miltiorrhiza EST-SSR molecular mark. The method mainly comprises the following steps of obtaining and pre-processing a salvia miltiorrhiza EST sequence, screening a SSR locus in the salvia miltiorrhiza EST sequence, designing and synthesizing the salvia miltiorrhiza EST-SSR primer, screening the salvia miltiorrhiza EST-SSR primer and the like. The method excavates the SSR locus aiming at the salvia miltiorrhiza EST and designs the PCR primer according to the flanking sequence of the SSR locus, and on the basis of this, the salvia specificEST-SSR mark system is established; and the method lays a solid foundation for genetic diversity evaluation, genetic relationship or idioplasmatic identification, genetic linkage description, molecular mark auxiliary selection, gene mapping cloning, functional genome analysis, related researches and the like of related species of the salvia miltiorrhiza and the salvia. The invention also discloses the EST-SSR locus specific primer obtained by the method for preparing the salvia miltiorrhiza EST-SSR molecular mark and the application thereof.

The invention relates to the technical field of gene engineering, and discloses a recombinant yeast strain, a construction method thereof and the application thereof. In the recombinant yeast strain, gal1, gal7, gal10 and ypl062w genes are knocked out, and the recombinant yeast strain comprises 6 gene segments which are integrated to the genome through yeast homologous recombination. On the above basis, the recombinant yeast strain is further integrated onto one gene segment of a yeast genome, so that a better recombinant yeast is obtained. According to the technical scheme of the invention, the yeast strain is constructed based on gene knock-out, and an optimized host cell for producing lycopene is provided. Meanwhile, functional genes crtE crtB and crtI of a specific source combination and used for the synthesis of lycopene, specific yeast endogenous genes and specific exogenous genes are selected. The above selected genes are integrated to the gene knock-out genome of the yeast strain, and then a brand-new recombinant strain capable of producing high-yield lycopene is obtained.

The invention discloses a method for performing transient expression by introducing a foreign gene into a poplar bioplast. The method comprises the following steps of: introducing a transient expression vector containing a foreign gene into a poplar bioplast by using PEG (Polyethylene Glycol)-Ca<2+>; culturing; and expressing the foreign gene in the bioplast. A poplar bioplast transient expression system constructed with the method is easy to operate and realize, a place with broad prospects is opened up for the research of functional genes of a poplar, and the transformation efficiency of the method can be up to 70 percent.

The invention discloses a screening method of Pacific oyster EST micro satellite label, which comprises the following steps: finding micro satellite site with Pacific oyster EST sequence announced by GeneBank and micro satellite searching software SSRhunter; screening and separating 2-6 base repeat unit micro satellite fragment (repeat times >5); getting repeat ESTs sequence with micro satellite; designing primer at two sides flank sequence of micro satellite repeat sequence; detecting optimization primer as the micro satellite label further; proceeding overall merit for micro satellite repeatability, stability, polymorphism; getting micro satellite label. The invention can be used for Pacific oyster species resource and genetic heterogeneity, molecular population genetics, species resource determination, genetic atlas construct, functional gene research, Pacific oyster breeding and cultivation.

The invention discloses a tobacco genome molecular marker probe and sequence collective group as well as an acquiring method and an application thereof. The probe and primer set has any nucleotide sequence as shown in all or partial No9-25. The new probe and primer sequence sets are from a series of new minitype reverse repetitive transposable elements separated and identified by the inventor of the invention. The probe and primer are acquired through a sequence group and independent sequence and through new sequence selection and identification, and can be applied to the following fields: 1, tobacco molecular marker auxiliary breeding, and selecting and breeding of new breeds of traditional tobacco to fasten the process; 2, construction of tobacco genetic spectrum and identification of tobacco DNA fingerprint; 3, important functional gene positioning and cloning in tobacco genome; 4, early-stage identification and selection of important agronomic character of tobacco; 5, genetic relationship identification of different tobacco breeds or strain materials; and 6, true-false identification of tobacco products (including cigarettes).

The invention discloses a method for constructing a kidney cell line of scophthalmus maximus, which comprises the following steps of: preparing a cell culture fluid first, taking a kidney tissue of the scophthalmus maximus as a material, using 0.5 percent trypsinase to digest the kidney tissue for 30 minutes at room temperature after the rinsing and shearing, adopting an 200-mesh nylon gauze to filter and collect cells, split charging the cells with the quantity of 5-10*10 counts / bottle into culture bottles with the growth area of 25 cm, and placing the culture bottles at a temperature of 24 DEG C for culturing; replacing the cell culture fluid with half quantity each four days, and using 0.25 percent trypsinase digest the cells for passage when primary cells grow to be monolayered; and performing passage once each 8 to 10 days, wherein the content of serum in the cell culture fluid is reduced to 10 percent from 20 percent when the 8th generation is reached, and the cell line is successfully established at the moment. The kidney cell line of the scophthalmus maximus obtained by utilizing the method is fibroid and can be continuously transferred for more than 40 generations; the cell line can be directly applied to pathogeny characteristics research, vaccine development and functional gene research; and the construction method is suitable to construct kidney cell lines of other fishes.

The present invention provides algorithm-based molecular assays that involve measurement of expression levels of genes, or their co-expressed genes, from a biological sample obtained from a prostate cancer patient. The genes may be grouped into functional gene subsets for calculating a quantitative score useful to predict a likelihood of a clinical outcome for a prostate cancer patient.

The invention discloses a larimichthys crocea liver cell line and an establishment method thereof. The larimichthys crocea liver cell line is preserved in the China center for type culture collection (CCTCC) On October 31, 2013 and has a preservation number of CCTCC NO: C2013157. The larimichthys crocea liver cell line can realize serial passage, can provide a large amount of larimichthys crocea liver cell lines, can be used for pathogenic characteristic research, vaccine development, functional gene research and breeding research, has excellent properties, comprises fibroblast-like cells and mainly comprises cells having parapodiums stretching out and irregular shapes. The larimichthys crocea liver cell line has the advantages of exuberant division, short passage time, easy digestion, good adherence rate and hunger resistance. The establishment method has good repeatability. The larimichthys crocea liver cell line has good stability.

The invention provides a product gene modeling method based on a behavior semantic-network knowledge model, which comprises the following steps: (1) a behavior semantic model of the product is built; (2) a mapping rule between the behavior semantic model of the product and a behavior semantic predicate is built; (3) a frame description proposal of the functional character and the geometrical characteristic of the product according to the mapping rule between behavior semantic model and the behavior semantic predicate; (4) the frame description proposal described in the step (3) is transformed into a corresponding gene code by applying the evolutionary design and the gene code technology, and a functional gene model of the product is built. Based on the code manner, the built functional model establishes a good basis for the evolutionary design and the automated innovation of the product, and fully prepares for further researches in the future.

The invention relates to the boron absorbing material and its preparation method and belongs to the multiple materials preparation and absorbing separation fields. It is to load multi-hydroxyl functional genes on the surface of inorganic multi-hole carrier in non-medium water to prepare the boron absorbing material with special structure and surface chemical character. The invention is suitable for the absorbing separation and collection of various enhydrous agents with boron and provides an effective way for the utilization of the boron resources and the treatment of the boron waster water.

The new quantitative character gene site locating method based on genome exon chip is to utilize exon sequence as one kind of molecular mark and utilize the detected data of human, rice and other allgenome or exon sequence chip of partial chromosome as colony locating molecular mark data for gene location via statistic location method. The present invention can obtain located functional gene sequence directly, and is suitable for all the quantitative character and complicated character of life. The research colonies include plant DH colony, F2 colony, near isogenic line, near isogenic introduction line, recombinant idiogamy line, animal family, etc.