195results about How to "Short screening cycle" patented technology

The invention provides fowlpox virus vector shuttle plasmid pTGP3 which comprises recombinant arms TKL and TKR, a bidirectional promoter PE/L, a fluorescent protein expression cassette, and a resistant marker gene and replication origin ori; the upstream and the downstream of the bidirectional promoter PE/L are respectively provided with cloning sites MCSL and MCSR; and both ends of the fluorescent protein expression cassette are provided with loxp sequences. The plasmid of the invention has two different screening markers, and the recombinant fowlpox virus prepared with the plasmid can express 1 to 3 types of gene with different meshes in the whole processes of the early and the later periods; the strong composite promoter with expression activity in the early and the later periods is applied so as to realize the all-process high-efficiency expression of a target gene; and the loxp sequences are introduced into both ends of the fluorescent protein expression cassette, so as to knock out the exogenous recombinant fowlpox virus screening markers. The invention lays foundation for the series and the scale application of the recombinant fowlpox virus in vaccine and biological drug research and development fields.

The Chinese medicine developing process includes selecting traditional Chinese recipe with determined curative effect, separating the components, optimizing the component combination, determining thescreening model and performing medicine effect test based on Chinese medicinal theory and compound Chinese medicine features, computer aided analysis to recognize effective components, combining modeand medicinal effect, and optimizing new Chinese medicine. The said process can raise the screening efficiency of effective components, shorten screening period and embody compound Chinese medicine features. Altering the combination mode of medical herb pieces into combination of effective components develops modern Chinese medicine preparation with clear matter basis and controllable quality, being safe and effective and coinciding with international standard.

The invention discloses an oligonucleotide library classification and assessment method based on capillary zone electrophoresis, and belongs to the field of creature isolation analysis. The method comprises the following steps of: step one, carrying out capillary zone electrophoresis on an oligonucleotide library, and obtaining a secondary library within an oligonucleotide library electrophoresis time range according to transfer time slicing collection; and step two, respectively mixing each secondary library with a homogeneous target molecule, carrying out capillary zone electrophoresis, and comparing the strong or weak of each secondary library and target molecule interaction, thus obtaining the strongest secondary library of the target molecule combining capacity. The classification and assessment method can be used for realizing the fractionation of complicated constituent oligonucleotide library, and obtaining the strongest secondary library of the target molecule combining capacity; and the strongest secondary library of the combining capacity is utilized as a next CE (capillary electrophoresis)-SELEX(systematic evolution of ligands by exponential enrichment) technical screening library, the screening range is reduced, and the screening period of an adaptation body is shortened.

The invention relates to a method for screening single chain antibodies of Microcystin-LR and verification thereof. The method comprises the following steps: carrying out two rounds of affinity screening on the biotinylated Microcystin-LR in a human source synthetic antibody library by using an avidin labeled magnetic bead and a negative screening method; extracting total plasmid DNA from phage colonies produced in the second round, carrying out enzyme digestion with enzyme Sfi I, recycling gel to obtain single chain antibody genes, connecting the single chain antibody genes with soluble expression carrier pAK100CL which is processed by enzyme digestion in the same way, and electrically transforming the connected carrier into colibacillus XL1-Blue to obtain soluble expression single chain antibodies; and verifying the soluble expression single chain antibodies by using a competitive time-resolved fluorescence immune analytical method. The invention has the advantage of quick, simple and convenient screening, and can well expose the Microcystin-LR three-dimensional structure into the incubation system. The verification on the screening result has the advantages of high detection signal and strong anti-interference capacity against stroma.

The invention relates to a friction wearing part test apparatus for a connecting rod small end bushing of an engine. The apparatus comprises a body (1), a bearing cover (2), a crankshaft (3), a connecting rod (4), a cylinder sleeve (5), a test piston (6), a piston pin (7) and a small end bushing (8). The crankshaft (3) performs rotary motion in a bearing structure composed of the body (1) and the bearing cover (2). The crankshaft (3) and the connecting rod (4) are connected, the connecting rod (4) is connected with the test piston (6) through the piston pin (7), the contact portion of the connecting rod (4) and the piston pin (7) is provided with the small end bushing (8), and the rotary motion of the crankshaft (3) drives the test piston (8) to perform up-and-down motion in the cylinder sleeve (5). The apparatus further comprises a support rod (9), a spring limiting disc (10), a motion disc (11) and a spring set (12). Compared to a whole machine test, the apparatus can reduce the cost for endurance testing of a connecting rod small end bushing.

The invention belongs to the technical field of cell research, and particularly relates to a method for constructing a laryngeal cancer cell line for stably expressing the green fluorescent protein. The method comprises the following steps: (1) designing and synthesizing a primer; (2) digesting the gene sequence of pLenti-puro carrier plasmids with BamHI and XhoI according to the gene sequence andpLenti-puro sequence of the green fluorescent protein, and connecting digested products to obtain a target carrier pLenti-puro-GFP; (3) verifying the carrier pLenti-puro-GFP with endotoxin extraction, digesting the pLenti-puro-GFP plasmids with BamHI and XhoI, and detecting the gene sequence by agarose gel electrophoresis, and sequencing the carrier pLenti-puro-GFP plasmids to detect the sequencesituation of the GFP-LC3; and (4) co-transfecting a 293T cell with the carrier pLenti-puro-GFP plasmids and a lentivirus package carrier, collecting virus supernatant, and transfecting a laryngeal cancer cell Hep-2 with the virus supernatant, infecting for 24 hours, then adding a culture medium containing puromycin, screening the cell, and observing under high content until a stable strain is obtained.

The invention relates to Saccharomyces cerevisiae and a preparation method thereof. A preparation method of high-nitrogen fresh yeast is mainly characterized by comprising the following steps of: (1) preparing a strain, i.e., Saccharomyces cerevisiae which is collected in the China General Microbiological Culture Collection Center with the collection number of CGMCC No.5004; (2) preparing a culture medium in a certain mass ratio; (3) pretreating sweet sorghum juice; (4) fermenting; (5) undergoing an adaptive phase at the ventilation quantity of 0.8-1.2 V.V.m and the stirring speed of 300-500 r/min for 4-6 hours, and adding nutritional salts, including 0.2-3 percent of (NH4)2SO4, 0.03-1.5 percent of MgSO4 and 0.2-1.5 percent of KH2PO4; (6) undergoing an exponential growth phase for 10-20 hours, fermenting at the fermenting temperature of 32-35 DEG C, undergoing a decline phase at the ventilation quantity of 0.3-0.5 V.V.m and the stirring speed of 80-150 r/min, stopping feeding liquid glucose, measuring the concentration of residual sugar in fermented cheese liquid, and stopping fermenting if the concentration of residual sugar is less than 0.5 percent; and (7) treating a fermentation liquid. In the method, the sweet sorghum juice is taken as a carbon source in the fermentation liquid. Compared with other carbon source culture mediums such as honey and beet, the sweet sorghum juice has the advantages of simple pretreating process, saving in energy consumption and increase in production efficiency.

The invention relates to a screening method of a cell strain of a GS expression system. The method includes the steps of: pressurizing methionine sulphoximine, selecting a cell pool being 25-35% in cell activity for performing monoclonal screening, and adding the cell pool to a semi-solid culture medium containing 10% of a conditional culture medium to form mono-clone (wherein final cell concentration is 150 cells/ml), and then performing screening and amplified cultivation to obtain a high-expression cell strain through a high-throughput cell screening system, and performing stability evaluation to the screened high-expression cell strain. The method can be used for obtaining the cell strain being more than 90% in stability.

The invention discloses a method for high throughput screening of microorganisms for preventing plant soil-borne fungal diseases, and belongs to the technical field of agricultural microorganisms. Themethod includes preparing the diseased oil with a high-concentration wheat root rot pathogenic bacteria culture and the soil, sowing germinated wheat seeds into the soil containing pathogenic bacteria to simulate the natural growth conditions and environment of plants and the pathogenic bacteria in the soil to the maximum extent. The method for screening for a pathogen-plant pathogenesis system of antagonistic microorganisms of the wheat bipolaris sp, the dense planting of the wheat reduces the space occupied by the screening, and the symptoms of the wheat root rot appear early, which reducesthe screening time, so that the high-throughput screening can be achieved. The screening efficiency of the microorganisms with better inhibition of pathogenic bacteria obtained in the early screeningis greatly improved, and a large number of microorganisms that cannot exert control effects in practical applications can be removed, so that the pace of microorganism screening, microorganism pesticide development and microorganism fertilizer research and development can be accelerated.

The invention discloses a method for screening myocardial ischemia resisting pharmacodynamic material basis of yang exhaustion treating decoction. The method comprises the following steps: 1, determining myocardial ischemia resisting endogenous metabolin of the yang exhaustion treating decoction and measuring the peak area; 2, determining in-vivo migration constituents in the yang exhaustion treating decoction and measuring the peak area; 3, performing correlation analysis on the peak area of the myocardial ischemia resisting endogenous metabolin of the yang exhaustion treating decoction and the peak area of the in-vivo migration constituents in the yang exhaustion treating decoction and screening out myocardial ischemia resisting pharmacodynamic material basis of the yang exhaustion treating decoction. The method disclosed by the invention is based on isoproterenol-induced rat myocardial ischemia model; an ultrahigh performance liquid chromatography-quadrupole rod-flight time mass spectrum non-targeted metabonomic technology and a targeted metabonomic technology based on an ultrahigh performance liquid chromatography-triple quadrupole rod mass spectrum are utilized to screen out 12 kinds of myocardial ischemia resisting pharmacodynamic material basis of the yang exhaustion treating decoction; thus, accuracy of the myocardial ischemia resisting pharmacodynamic material basis ofthe yang exhaustion treating decoction is improved, a screening period is shortened, and screening consumption cost is reduced.

The invention discloses a method for screening root media by using tobacco sterile seedlings. The method comprises the following steps: A, sterilizing tobacco seeds; B, cultivating the sterile seedlings; C, shearing the sterile seedlings; D, allocating a plurality of root media to be screened; and E, screening the optimal root media. The method is simple in process, labor-saving and time-saving, economic and reliable, and can shorten the screening period effectively, simplify the screening program, and save the screening material; and the sterile seedlings materials are obtained by seeds in a culture vessel directly, and enter the stage of root screening directly, which omits the trouble of cultivating adventitious buds as compared with the conventional screening test, so that manpower, material resources and time are saved. The tobacco sterile seedlings are adopted to screen the media, and the tobacco sterile seedlings screening can synchronize with the callus induction, so that the test schedule is speeded up. Besides, the induction effect of the method is consistent with that of a method for screening root media by using adventitious buds.

The invention relates to the field of biomedicine, in particular to a preparation method of snake venom thrombin-like protein. Firstly, the venom tissue of Agkistrodon akistrosus is isolated, and a phage display library is constructed. Using fibrin/fibrinogen as a substrate, three rounds of panning selected, a new thrombin-like gene from Agkistrodon venom was obtained. The present invention also constructs the prokaryotic expression vector pET-32(a)-TLE of the viper venom thrombin gene of Agkistrodon akistrodon, and obtains the recombinant snake venom thrombin with biological activity after induced expression, affinity purification and molecular sieve purification protein. The enzyme can be used to prepare cardiovascular thrombolytic drugs.