724 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.

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 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 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.

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 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.