474 results about "High-throughput screening" patented technology

The embodiment of the invention provides a method for obtaining a tyrosine ammonialyase mutant based on high-throughput screening, the tyrosine ammonialyase mutant and application of the tyrosine ammonialyase mutant. The mutant is subjected to any one of the following amino acid mutations on an amino acid sequence as shown in SEQ ID NO.2: A245G + A247T, and D192V + A245G + A247T. According to the embodiment of the invention, an amino acid sequence as shown in SEQ ID NO.2 is taken as a template (basis), a key active site of the template is subjected to mutation transformation, a mutant library is constructed, and a series of mutants with remarkably improved catalytic activity and / or conversion rate are screened; the tyrosine ammonia lyase mutant provided by the embodiment of the invention can selectively catalyze L-tyrosine to generate p-coumaric acid, and has excellent yield and / or higher yield, which indicates that the mutant has more significant application prospects in catalysis of L-tyrosine to generate p-coumaric acid.

The invention belongs to the technical field of antibacterial peptide activity identification and evaluation, and relates to an antibacterial peptide activity and MIC value combined prediction framework based on cross-modal deep learning, and the framework uses a protein language model ESM2 to respectively carry out token-level semantic embedding coding on an antibacterial peptide sequence and a pathogen protein sequence; performing cross-modal feature extraction and fusion through a multi-branch structure comprising a Mama module, a multi-head self-attention mechanism and DASM 1D convolution; a multi-task decoding structure is adopted to realize antibacterial peptide activity classification and MIC value regression prediction at the same time; according to the method, functional characteristics in the sequence can be effectively mined, the accuracy and generalization ability of antibacterial peptide activity and MIC value prediction are remarkably improved, and a reliable calculation tool is provided for high-throughput screening and rational design of the antibacterial peptide.

The invention discloses a 3-sterone-delta1-dehydrogenase mutant and application of the 3-sterone-delta1-dehydrogenase mutant in preparation of a steroid drug intermediate. The mutant is obtained by performing single mutation or multiple mutation on 551, 468, 48 and 157 sites of an amino acid sequence of 3-sterone-delta1-dehydrogenase as shown in SEQ ID NO: 1. The 3-ketosterone-delta1-dehydrogenase is subjected to molecular modification through directed evolution and semi-rational design, the mutant with improved enzyme activity is screened in a high-throughput mode, the mutant can effectively improve the conversion rate of a substrate 11alpha, 17alpha-dihydroxyprogesterone, and the substrate conversion rate of the mutant ISM-2 is improved to 92.4% compared with 36.7% before mutation.

The invention belongs to the field of biological pharmacy, and discloses a GPCR-based ultramicro drug screening method and application thereof in screening of antipruritic traditional Chinese medicines. According to the invention, an Echo ultrasonic pipetting system is combined with a GPCR function experiment (beta-arrest recruitment) to establish the ultramicro screening method for screening the GPCR ligand. The Echo 550 system utilizes ultrasonic waves to beat liquid into 2.5 nL liquid drops and then transfer the liquid drops into a 384 or 1536 pore plate, rapid and accurate sample adding of at least 2.5 nL (1 / 400 mu L) can be achieved, ultra-micro screening and high-throughput screening can be achieved at the same time, and the method is particularly suitable for screening of itching relieving drugs.

The invention relates to a high-throughput screening method and system for a self-assembled single-layer thermal interface material and a medium. The high-throughput screening method comprises the following steps: constructing an interface heat transport simulation system based on a self-assembled monomolecular layer; screening set preparation is carried out on the end group structure, and an end group screening set and a pre-calculation set are constructed; carrying out molecular dynamics simulation on the self-assembled monomolecular layer corresponding to the pre-calculation set to obtain a physical association index; training a machine learning model by taking the physical association index as a machine learning objective function; carrying out machine learning prediction on the uncalculated screening candidate data set based on the trained machine learning model, and carrying out parallel screening of physical association indexes to obtain a simulation verification set; and carrying out molecular dynamics simulation for verification. Compared with the prior art, high-throughput screening of the self-assembled single-layer thermal interface material is achieved, and the method has important significance on solid-liquid interface heat conduction characteristic regulation and control design under data driving.

The invention relates to the technical field of molecular biology, in particular to a multichannel virus reporting system based on TAT trans-activation. The invention relates to a multichannel virus reporting system based on TAT trans-activation. The multichannel virus reporting system comprises an activated virus and a reporting cell, the activated virus is a human enterovirus A71 for expressing TAT protein; the reporter cells are cells transfected with reporter plasmids; the reporter plasmid comprises an LTR sequence and a plurality of reporter genes which are started to express by the LTR sequence. When the virus is activated to infect the reporter cell, the TAT protein carried by the virus enters the reporter cell to activate the expression of the downstream reporter gene, and the expression level of the reporter gene can reflect the virus replication condition, so that the screening or mechanism research of related antiviral drugs can be realized. According to the technical scheme, the technical problem that the flexibility, stability and high-throughput screening adaptability of a virus reporting system in the prior art are not ideal can be solved, and the virus reporting system has ideal popularization and application prospects.

The invention discloses a method for improving the synthesis level of L-tryptophan in escherichia coli based on ARTP mutagenesis high-throughput screening, and belongs to the technical field of bioengineering. According to the invention, the L-tryptophan biosensor pSensor-trp3 is constructed, and the L-tryptophan biosensor pSensor-trp3 is good in response under the condition that the exogenous addition concentration range of L-tryptophan is 0-300mg. L <-1 >. High-throughput screening is performed by using a flow cytometry, a dominant mutant strain YB-2 is obtained through iterative mutagenesis, and the shake flask L-tryptophan yield and the sugar-acid conversion rate of the dominant mutant strain YB-2 are 5.68 g.L <-1 > and 0.103 g.g <-1 > respectively and are increased by 11.4% and 10.7%. Through second-generation whole genome re-sequencing comparative analysis, it is speculated that the genes ynfB and waaO possibly have a promoting effect on synthesis of L-tryptophan. The L-tryptophan biosensor used in the invention also provides reference for high-throughput screening and transformation of other related L-tryptophan production strains.

An automated high throughput screening platform for identification of compounds and genomic constructs having mitochondrial function modulating activity and compounds so identified are provided. Use of such compounds for modulating mitochondrial function are also disclosed. A high-throughput screening method of determining genetic mechanisms of action for therapeutic or toxic agents in genetic or pharmacologic models of disease are also provided.

The invention belongs to the field of biotechnology and environmental governance, and particularly relates to a manganese peroxidase mutant based on high-throughput screening modification and application of the manganese peroxidase mutant to straw degradation. Aiming at the problems of low natural secretion amount, insufficient specific activity, complex preparation process and the like of the existing manganese peroxidase, the method comprises the following steps: firstly, constructing engineering bacteria based on an escherichia coli exocrine system, constructing a mutant library through error-prone PCR in combination with a directed evolution technology, and performing high-throughput screening by utilizing an ABTS colorimetric method; the recombinant manganese peroxidase mutant with high catalytic activity is obtained, and the enzyme activity is improved by 1.29 times compared with that of a wild type. Furthermore, recombinant engineering bacteria are constructed on the basis of a bacillus subtilis exocrine system, and efficient secretion of the mutant enzyme is achieved. The mutant enzyme preparation is used for wheat straw degradation, and the lignin degradation rate reaches 41.37% (improved by 30.09% compared with that of a wild type). The method is simple and convenient in process, low in cost and remarkable in effect, and has a wide industrial application prospect.

The invention discloses a simulated screening and process optimization method of an anhydrous hydrogen fluoride desulfurization adsorbent, and relates to the technical field of anhydrous hydrogen fluoride, and the method comprises the following steps: constructing a database, and establishing a basic screening library; constructing an intelligent screening engine, and performing force field parameter correction on the materials in the basic screening library to capture a high-polarity solvent effect; calculating and extracting adsorption selectivity, Henry coefficients and working capacity indexes, and screening out a primary candidate material set; a core candidate material is optimized; determining an optimal adsorbent model; the microscopic adsorption data of the optimal adsorbent model are converted into macroscopic process parameters, the macroscopic process parameters are imported into a macroscopic process simulator for prediction, and process optimization is completed. According to the method, the strong polar solvent effect is simulated by constructing the corrected force field environment, and high-throughput screening of the adsorption materials is achieved in combination with an active learning strategy; an electronic grade refinement and modification strategy is utilized to optimize adsorption sites, and microcosmic adsorption data and a macroscopic mass transfer model are coupled to realize accurate prediction and optimization of an adsorption process.

The invention discloses a method for screening an MOFs material with excellent CH4 adsorption separation performance based on high-throughput calculation, a method for improving the screening efficiency and a preparation method. The screening method comprises the following steps: constructing a screening database of MOFs material structure information; calculating physical and chemical characteristic parameters by utilizing computational chemistry and material simulation software, and further setting a screening standard aiming at CH4 adsorption; carrying out preliminary screening through a high-throughput screening technology; predicting the competitive adsorption behavior of the primarily selected material in the mixed gas by means of a molecular simulation technology, and performing secondary screening; and finally, determining the final MOFs material in combination with an actual application environment. According to the method, the high-performance MOFs material is rapidly and accurately screened out from a large number of materials, the research and development period is effectively shortened, the development cost is reduced, an efficient and feasible solution is provided for resource utilization of ultralow-concentration CH4 in coal mine windblown gas, and energy utilization and environmental protection are assisted.

By engineering genetic traits associated with photosynthesis in plants, transgenic plants with increased crop yield can be generated, which will ultimately lead to increased food production in the field. To obtain beneficial genetic traits for enhanced photosynthesis, accelerated evolution is applied under salt stress in a cyanobacteria species, Synechococcus elongatus PCC 7942, by conditionally suppressing its mismatch repair system. Through high-throughput screening for biomass accumulation and sucrose production, it is discovered that specific amino acid changes in the pre-Dl protein of Photosystem II (PSII), encoded by the psbA genes, can improve efficiency of photosynthesis under salt stress, as well as pathogen resistance. Next, the same amino acid changes were made in Arabadopsis and proved that photosynthetic efficiency (measured via growth rate) is also increased in this model vascular plant.

A well plate can include a plastic component having an array of microwells, each defining a respective opening sized to receive and contain a volume of fluid. A printed circuit board (PCB) material can be arranged against and accessible through the microwell openings, allowing fluid contact with the PCB material at a first side. An array of integrated circuits (ICs) can be individually positioned to correspond with respective microwells and electrically interconnected via conductive interconnections within the PCB material, such as to facilitate electronic assays within standardized well plates for high-throughput screening applications.

The invention provides a method for high-throughput rapid screening of food fermentation strains. The method comprises the following steps: mixing a to-be-detected bacterial sample with a culture medium without a yeast product, injecting an obtained mixed bacterial liquid and an oil phase into a liquid drop generation chip, generating liquid drops, and collecting the liquid drops for culture; injecting the cultured liquid drops, the oil phase and SYTOTM16 fluorescent dye liquid into a liquid drop micro-injection chip, collecting the generated liquid drops, injecting the liquid drops and the oil phase into a micro-fluidic sorting chip, carrying out fluorescence detection, collecting target liquid drops, and carrying out demulsification treatment and coating culture. The method can realize rapid and accurate high-throughput screening of target strains, and especially can directly quantify the growth activity of microorganisms under the stress of acid and salt, thereby realizing efficient and accurate screening of acid-resistant and salt-resistant strains. Moreover, the method does not need to carry out gene modification on the strain, avoids the problems of biological safety and technical complexity possibly caused by gene manipulation, retains the natural characteristics of the strain, is suitable for large-scale high-throughput bacterium screening, and has high application value.

The invention belongs to the field of microbial breeding and fermentation engineering, and particularly relates to a saccharomyces cerevisiae mutagenesis strain with high yield of S-adenosyl-L-methionine as well as a mutagenesis method and application of the saccharomyces cerevisiae mutagenesis strain. Saccharomyces cerevisiae is induced to generate genetic diversity through multiple rounds of ultraviolet mutagenesis, and high-throughput screening is performed in combination with strain color phenotypic difference; and the mutant strain HY2402-Z18 with high yield of S-adenosine-L-methionine and genetic stability is obtained. The SAM yield of the mutagenic strain is increased by 40.1% compared with that of an original strain, the SAM yield reaches 13.44 g / L when the mutagenic strain is fermented for 60 h in a 5 L fermentation tank system, the unit yield reaches 0.156 g / L / OD, and the methionine conversion rate is 75%. After thalli are collected through centrifugation of fermentation liquor, an S-adenosine-L-methionine product with the purity of 98%-100% can be obtained through purification of ion exchange resin, no by-products are accumulated, the yield, the substrate conversion rate and the production efficiency of the S-adenosine-L-methionine can be remarkably improved when the mutagenesis strain is used for fermentation production, and the production cost is reduced. The production cost is effectively reduced, the product purity and quality are improved, and the method is suitable for industrial large-scale production of SAM.

The invention discloses an esterification reaction lipase screening method based on covalent bond docking, and belongs to the technical field of protein docking simulation. According to the method, parameters of an acylase intermediate are constructed, and then covalent docking and energy screening are performed on the acylase intermediate, so that prediction of lipase reaction activity and substrate selectivity is realized, and the defects that carbonyl carbon cannot be automatically identified, Ser-C covalent bonds cannot be established, double bonds / bond angles cannot be managed, and the reaction efficiency is low in traditional docking are overcome. The problems that in a covalent system, simulation of a nucleophilic attack path lacks a unified distance, angle and dihedral angle constraint system, and automatic and high-throughput screening of multiple lipases and multiple substrates is difficult to achieve are solved.

The invention relates to the technical field of biological detection, and discloses a caenorhabditis elegans reproductive toxicity intelligent evaluation system and an application method.The caenorhabditis elegans reproductive toxicity intelligent evaluation system comprises a standardized exposure and imaging module used for executing a high-flux pollutant exposure experiment under a controlled environment condition and synchronously collecting nematode behavior and gonad image data; and the multi-modal data acquisition unit is connected to the standardized exposure and imaging module and is used for extracting and preprocessing behavioral parameters and reproductive toxicity indexes from the acquired images. According to the method, 12 behavior parameters and 8 core reproductive indexes are automatically extracted by constructing a multi-modal data acquisition unit and utilizing an improved U-Net model and a specific algorithm, so that full-process automation from image acquisition to key index quantification is realized. The problems of low efficiency, large subjective deviation and poor repeatability caused by dependence on manual interpretation in the prior art are solved, and the objectivity of the evaluation result and the high-throughput screening efficiency are remarkably improved.

The invention relates to a gene expression cassette and application thereof. The gene expression cassette comprises a tyrosinase gene, a transcriptional repressor protein gene, a melanin response promoter and a reporter gene, and the reporter gene is located at the downstream of the melanin response promoter and is regulated by the promoter. The gene expression cassette with a specific structure is designed, melanin in cells can be responded, a melanin biosensor can be further constructed by utilizing the gene expression cassette, the gene expression cassette has good specificity, and high-throughput screening of strains can be realized. Furthermore, ALE evolution is carried out on the strain for producing melanin, high-throughput screening is carried out based on a designed sensor, the strain for producing melanin with high yield is obtained, through shake-flask culture and 5L fermentation tank culture, the yield of melanin reaches up to 10.84 g / L and 29.04 g / L respectively, and an efficient and feasible solution is provided for large-scale production of melanin.

The invention relates to a thermal analysis method and a measuring device for a solidification process after modification treatment of aluminum and aluminum alloy, and the thermal analysis method comprises the following steps: adding seed crystal particles into an overheated melt in a vacuum environment or an inert atmosphere through a feeding device; cooling and solidifying the overheated melt, and collecting and recording a cooling curve T-t of the seed crystal particles through a high-speed temperature collecting device; the cooling curve is subjected to derivation calculation, and thermal characteristic parameters in the melt solidification process are represented; comparing the thermal characteristic parameters, and judging the deterioration effect. According to the method, the seed crystal particles are placed into the melt, melt solidification data are collected, and efficient judgment and high-throughput screening of the seed crystal particles and the second-phase modified refined seed crystal particles in the aluminum alloy are achieved by means of the quantitative relation among the seed crystal particle characteristics, the local temperature and the supercooling degree.

The invention discloses a urate oxidase mutant and application thereof in improving the thermal stability of urate oxidase, and belongs to the technical field of biology. The urate oxidase mutant disclosed by the invention is T68L / T75S / E222D / K299E, T68L / T75S / K299E or T75S / K299E, and the urate oxidase mutant disclosed by the invention is T68L / T75S / K299E. According to the method, the key sites influencing the thermal stability are mined in the modes of directed evolution, high-throughput screening, multiple virtual screening and the like, and the stability is improved through mutation. And finally, a plurality of key sites and mutants influencing the stability are excavated, and the thermal stability is improved.

The invention discloses a method for microfluidic high-throughput screening of AKK bacteria. The method comprises the following steps: 1) preparing a microfluidic chip; 2) collecting fresh excrement of healthy volunteers, diluting, and adding a fluorescence labeled antibody to obtain a dispersion phase; and (3) preparing liquid drops by taking paraffin oil and Span-80 as a mobile phase, enabling the liquid drops to enter a liquid drop culture tank through an S-shaped liquid drop circulation channel, culturing, introducing the mobile phase, introducing the liquid drops in the liquid drop culture tank into a liquid drop collection tank, taking out, detecting a fluorescence signal, culturing the liquid drops with the fluorescence signal on the improved brain heart infusion broth solid culture medium, and obtaining the brain heart infusion broth. Obtaining a to-be-identified strain; 4, the strain to be identified serves as a template, PCR amplification is conducted, sequencing is conducted on an amplification product, BLAST sequence comparison is conducted on a sequencing result on an NCBI website, and the target strain AKK bacterium is screened out. The method has the advantages of being high in throughput, simple in experimental process and high in specificity, and the screening efficiency of the AKK bacterium can be improved.

The present disclosure relates generally to methods and systems of producing Schwann cells from pluripotent stem cells under fully defined conditions. The Schwann cells produced by the disclosed methods find applications as models of the enteric nervous system, tools for high throughput screening of potential therapeutics for treatment of enteric neuropathies, and in regenerative medicine.

The invention belongs to the field of neuroblastoma tissue analysis, and particularly discloses a child neuroblastoma tissue microarray chip and a preparation method thereof.The child neuroblastoma tissue microarray chip comprises a glass slide base material, and a plurality of neuroblastoma tissue samples arranged in a dot-matrix mode are arranged on the glass slide base material; by integrating a plurality of neuroblastoma samples including various pathological typing, grading and clinical staging types of neuroblastoma in a tissue block, efficient sample processing and analysis are achieved, comprehensiveness and representativeness of research are ensured, a large number of samples are analyzed simultaneously through a microarray technology, and the accuracy and accuracy of analysis are improved. The requirements for high-throughput screening of specific genes and expression proteins thereof in scientific research activities are met, the prediction capacity for tumor prognosis and the guidance accuracy for treatment schemes are improved, and neuroblastoma tissues are analyzed more efficiently and comprehensively.

The invention provides a high-throughput screening method for an AI energized beneficiation flotation reagent, and belongs to the technical field of mineral flotation reagent screening. According to the method, high-throughput screening of multiple candidate mineral flotation reagents is achieved through the flotation performance prediction model constructed through the artificial intelligence model, the research and development cycle and test cost of the mineral flotation reagents can be reduced, and the screening efficiency of the mineral flotation reagents can be improved.

The present invention discloses a plasmid vector and cell line that can induce targeted genomic DNA double-strand breaks. The present invention constructs a nuclease AscI expression plasmid based on a lentiviral vector backbone by integrating the Tet-on chemically inducible transcription control system and the ERT2 ligand-dependent nuclear localization control system. The vector regulates the transcription of AscI through the Tet-on system and uses the 4-hydroxytamoxifen-induced ERT2 system to achieve spatiotemporal specific nuclear translocation of the endonuclease AscI. The present invention also constructs and screens a cell line that stably expresses AscI. With the help of small chemical molecules, the spatiotemporal expression of the AscI endonuclease in cells can be precisely regulated, thereby inducing DNA double-strand breaks at specific sites in the genome. The present invention provides an efficient and accurate research tool for the field of DNA damage repair, which can be widely used in studying DNA damage repair mechanisms and high-throughput screening of related drugs.

The invention provides a method for high-throughput screening of an alpha-glucosidase inhibitor from boxthorn leaves based on a multi-shell MOF immobilized enzyme, and aims to overcome the bottlenecks of low throughput, tedious operation, high cost and the like of a traditional screening method. The core process of the method comprises the following steps: preparing a boxthorn leaf sample solution and pre-evaluating the in-vitro alpha-glucosidase inhibitory activity; a multi-shell MOF immobilized enzyme technology is coupled with a UPLC-QTOF-MS / MS technology, so that targeted screening, efficient separation and accurate identification of active components are realized; verifying the alpha-glucosidase inhibitory activity of the target compound; and predicting the binding site of the small molecule compound and the enzyme through a molecular docking technology. The method has the core advantages of high specificity, high sensitivity, high flux, repeated utilization of enzyme receptors and the like by depending on the inherent characteristics of an immobilized enzyme technology, can synchronously screen and identify various inhibitors, effectively integrates the resource advantages of a boxthorn leaf natural product and the high efficiency of a modern high-flux screening technology, and has a wide application prospect. And a key technical support is provided for diversified development of diabetes treatment medicines.

This invention belongs to the fields of molecular biology and bioinformatics, specifically relating to a truncated bovine virus-encoded protein phage library, its construction method, and applications. This library covers all 162 known bovine viruses that host bovine subfamily viruses, containing 51,794 non-redundant peptide coding sequences of 56 amino acids each. These peptides are fused and displayed on the surface of the T7 phage capsid, with all coding sequences having a uniform total length of 200 bp. The library construction was achieved through sequence acquisition, truncation and redundancy removal, codon optimization and DNA synthesis, and in vitro ligation and packaging with T7 phage. The resulting library exhibits a positivity rate of 93% and a titer of 2.8 × 10⁻⁶. 10 PFU / mL. This invention features an optimized single-round high-throughput screening process adapted to clinical bovine serum samples, enabling efficient identification of bovine virus B-cell epitopes and providing tools and candidate targets for bovine virus vaccine development and diagnostic reagent development.

The invention relates to a high-yield protein glutaminase Chryseobacterium prion strain and a preparation method thereof. The strain is obtained by carrying out ARTP and NTG compound mutagenesis treatment on an original strain of Chryseobacterium prion and carrying out high-throughput screening on the strain subjected to the compound mutagenesis treatment. The strain is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC M 20251792. The invention also relates to a method for preparing protein glutaminase by using the strain of the invention. The enzyme activity of the protein glutaminase of the strain is 6 times that of an original strain, and the production efficiency of the protein glutaminase can be remarkably improved.

The invention discloses an RNF130 gene responding to a BMAA signal, an encoded protein and application, and the RNF130 gene is found to be a gene strongly responding to the BMAA signal through high-throughput screening. After the RNF130 gene is knocked out, cell proliferation inhibition caused by BMAA is reduced for mammalian cells. In addition, the RNF 130 transduction the BMAA signal to the downstream GSK-3beta protein, thereby correlating with the Wnt signaling pathway. After the activity of GSK-3beta is inhibited by using a small-molecule GSK-3beta inhibitor, the reaction of RNF130 to a BMAA signal is changed, and the influence of cell proliferation inhibition caused by BMAA can be relieved by cells. According to the present invention, the cell proliferation inhibition caused by the BMAA can be relieved by the knockout of the RNF130 gene and the inhibition of the GSK-3beta activity, such that the key response protein and the molecular mechanism of the cell proliferation toxicity caused by the algal toxin BMAA are provided so as to provide the important reference target for the research and development of the antidote for preventing or treating the BMAA toxicity or the research and development of the drug aiming at the RNF130 related structural domain in the future.

The invention discloses a method for high-throughput screening of high-activity lactic acid bacteria based on multi-index fusion and application, and belongs to the field of lactic acid bacteria screening, the method comprises the steps of bacterial cell enrichment, single-cell suspension preparation, physiological status and activity evaluation of cells, flow cytometry analysis, cell sorting and strain identification; in the step of enrichment of the bacterial cells, a sample is activated and enriched by using an improved liquid culture medium and an improved solid culture medium, and activated and enriched bacterial suspension is obtained. The invention provides the method for detecting the activity of the fermentation product and the lactic acid bacteria stored for a long time, the method is convenient and rapid, the screening time is greatly shortened, and the screening efficiency is improved.