20results about How to "Improve expression efficiency" patented technology

This invention discloses a method for mRNA circularization and translation based on ribozyme catalysis. The method utilizes a Td ribozyme sequence to perform in vitro mRNA circularization, yielding circRNA; and introduces an IRES sequence into the circRNA. The IRES sequence is derived from MCDV, CVB3, HCV, PTV-1, RSV, or TSV. Through screening, this invention ultimately determined that the Td ribozyme sequence has the highest circularization efficiency, and that the IRES sequences derived from MCDV, CVB3, HCV, PTV-1, RSV, or TSV have strong ribosome binding capabilities. The MCDV IRES sequence is adaptable to various cellular environments. This invention is of great significance for improving the stability and expression efficiency of mRNA in vivo and promoting the development of circular mRNA technology.

The application discloses a method for increasing 2-pyrrolidone synthesis and belongs to the technical field of bioengineering. tacm The CoA transferase from Butyricicoccus faecihominis is expressed in the coryneform bacterium glutamicum, and the CoA transferase is secreted and expressed by means of P The promoter and the NS signal peptide strengthen the secretory expression of the CoA transferase, so that the constructed strain can ferment 2-pyrrolidone in a glucose carbon source and in a pH 7.0 environment, and the yield of the 2-pyrrolidone can reach 12.13 g / L in 96 h of flask fermentation.

The invention discloses an eurytopic chlamydomonas reinhardtii expression vector based on a PSAD promoter and a 3 * Flag tag as well as a construction method and application of the eurytopic chlamydomonas reinhardtii expression vector. The nucleotide sequence of the eurytopic chlamydomonas reinhardtii expression vector is shown as SEQ ID No.1; the vector comprises a PSAD promoter, a target gene insertion site EcoR V, a 3 * Flag tag at the 3'terminal, an rbcS2 terminator, a TUB2 promoter, a hygromycin B screening gene and an rbcS2 terminator. The construction method comprises the following steps: constructing a skeleton of a pHyg-PSAD-3Flag vector; constructing a sequence fragment fused with a 3 * Flag tag; and connecting the sequence fragment fused with the 3 * Flag tag with a plasmid skeleton fragment through T4 DNA (deoxyribonucleic acid) enzyme so as to construct the pHyg-PSAD-3Flag vector. According to the invention, the expression plasmid of the chlamydomonas pHyg-PSAD-3Flag can be used for efficiently screening algae strains capable of stably expressing target protein.

The invention belongs to the field of biological medicine, and particularly relates to a 5 'UTR sequence and application thereof. The technical problem to be solved by the invention is to improve the expression efficiency of mRNA drugs. The main technical scheme of the invention is as follows: two 5 'UTR sequences, which are SEQ ID NO: 1 and SEQ ID NO: 2, are provided; and a nucleic acid molecule comprising the 5 'UTR sequence. Compared with a 5 'UTR sequence optimized in the nature, the mRNA constructed by the 5' UTR has the advantages that the expression efficiency of a target gene is obviously improved; the preparation method is beneficial to reducing the administration dosage and administration frequency of the mRNA drug, reducing the cost of the mRNA drug and reducing the risk of adverse reaction after administration, and has a good application prospect in the industrial production of the mRNA drug.

The invention relates to a dialogue processing method and device, equipment, a storage medium and a program product. The method comprises the following steps: displaying a dialogue interface, and displaying first input content in the dialogue interface; displaying a first prompt content in the dialogue interface; the first prompt content comprises at least two pieces of non-text first visual content; each first visual content has an interaction attribute; the first visual content is used for supplementing the first input content; in response to a trigger operation on the first visual content, displaying a second input content in the dialogue interface; the second input content is obtained on the basis of triggering the selected first visual content. In this way, the user is more accurately and efficiently assisted to clarify the real information demand thereof.

The application discloses an anti-glyphosate EPSPS protein and application thereof. The amino acid sequence of the EPSPS protein coded by the gene is shown as SEQ ID NO. 2, the nucleotide sequence optimized for tobacco codon is shown as SEQ ID NO. 3, and the EPSPS protein is fused with a chloroplast signal peptide. The gene is introduced into Nicotiana benthamiana by an agrobacterium-mediated method, and the obtained transgenic plant shows extremely high glyphosate resistance and can tolerate a concentration of up to 36000 mg / L, which is significantly better than that of a traditional CP4-EPSPS transgenic plant. The gene is screened from microorganisms by using artificial intelligence assisted evolution analysis, structure prediction and semantic clustering, and has the characteristics of high-efficiency expression and strong resistance, and can be widely applied to herbicide-resistant breeding of various crops, thereby reducing production cost and environmental risk.

The invention relates to an anti-Theras gene derived from a cold spring, a screening method, an activity determination method and application of the anti-Theras gene, and particularly discloses an anti-Theras gene derived from the cold spring, and the nucleotide sequence of the anti-Theras gene is shown as SEQ ID NO: 1. According to the application, a novel anti-Theras gene is identified from a deep sea cold spring environment for the first time, and heterologous expression and plaque experiments prove that the novel anti-Theras gene can effectively antagonize a Theras defense system of a host. According to the anti-Theras gene provided by the invention, the types and sources of anti-defense genes are expanded, and a gene blank for non-classical defense system Theras from a cold spring environment is provided.

This invention provides a method for site-specific replacement of the creatine kinase promoter in the Huaxi bovine gene promoter, belonging to the field of animal gene editing and genetic breeding technology. It also provides a method for enhancing the function of bovine muscle cells. FTO Gene expression reagents include crRNA for upstream site knockout, crRNA for downstream site knockout, a Cas12i nuclease expression vector, and an FTO homologous substitution vector; the FTO homologous substitution vector includes... FTO upstream homologous arm of gene promoter, MCK gene promoter and FTO Downstream homologous arm of the gene promoter. The reagent was used in West China bovine cells. FTO Application in gene promoter replacement. This invention can specifically improve the expression efficiency of genes related to fat development in muscle tissue, providing an efficient and precise technical means for the genetic improvement of the performance of beef from Western China.

This invention belongs to the field of biomedical technology and discloses a chimeric antigen receptor targeting CD20, its gene expression construct, and its applications. The chimeric antigen receptor provided by this invention specifically includes an amino acid fragment with the sequence shown in SEQ ID NO:29 or a variant fragment having at least 85% sequence identity with SEQ ID NO:29. Its functional fragments, including the signal peptide, antigen-binding region, hinge region, transmembrane region, intracellular co-stimulatory domain, and signal transduction domain, form an organic whole, working synergistically to endow the chimeric antigen receptor with good CD20-targeting binding, T-cell activation, and cell-killing effects, demonstrating excellent application potential in the preparation of drugs for the treatment of relapsed / refractory B-cell malignancies.

This invention belongs to the field of genetic engineering technology and discloses a method for synthesizing mussel adhesive protein using gene recombination technology, as well as related recombinant vectors and strains. The method uses the core functional genes of mussel byssalin 3 and 5 as templates, and constructs a recombinant expression vector pET-20b-fp-cut through artificial synthesis and overlapping PCR gene tandem. This vector is then transformed into *E. coli* BL21 to obtain the recombinant strain BL21 / pET-20b-fp-cut. A gradient-scale fermentation process is used to optimize the efficient expression of the target protein. The product is then extracted using a salting-out-acetic acid extraction combined with tyrosinase-catalyzed purification process. Finally, multi-dimensional quality testing is used to verify the indicators. This invention also provides the aforementioned recombinant expression vector and recombinant strain. This method can realize the industrial production of mussel adhesive protein, with a fermentation unit of 5.2 g / L, a dopa content of 11.2%, and a purity of 96.5%. It also exhibits low immunogenicity and meets quality standards, showing broad application prospects in biomedicine, cosmetics, and other fields.

The present invention relates to an o / w emulsion comprising an oil phase, an aqueous phase and particles dispersed in the aqueous phase, wherein the particles are lipid nanoparticles (LNP) comprising a nucleic acid component, the particles being capable of stabilizing the oil-water interface. The invention also discloses a pharmaceutical composition, such as a vaccine, comprising the o / w emulsion.

The invention discloses an engineering information representation method, and relates to the technical field of engineering drawing, and the method comprises the steps: representing each engineering object in a target plane engineering drawing by employing a keyword in an engineering information keyword library, and obtaining the digital information of the target plane engineering drawing; the engineering information keyword library comprises keywords of various engineering objects, and the keywords of each engineering object comprise serial numbers, external dimension parameters, internal dimension parameters and position parameters; each engineering object has a unique number, the internal size parameter is internal structure size information of the engineering object, and the position parameter comprises at least one piece of position information of the engineering object; and storing the digitized information of the target plane engineering drawing. The method can improve the expression efficiency of the engineering information.

This invention provides a high-throughput and efficient method for screening multiple target genes that can regulate mRNA expression levels in cells by co-transfecting or sequentially transfecting mRNA with a gene silencing library targeting genes in cells using lipid nanoparticles. Several target genes that can regulate mRNA protein expression levels through the action of siRNA have been discovered. By manipulating these genes that regulate mRNA expression levels, the translational capacity of mRNA in mRNA vaccine and drug development can be improved, mRNA protein expression efficiency can be increased, and immunogenicity and toxicity can be reduced.

The application provides a repeated protein expression method and integrated device using anti-degradation circular DNA and circular RNA. The repeated protein expression method comprises a DNA template capable of producing circRNA in a cell-free protein expression system; the DNA template is a double-stranded DNA template with closed ends formed by two single-stranded DNAs; the double-stranded DNA template is connected with a closed linker at two ends of a linear DNA template; the closed linker comprises a 5' end linker and a 3' end linker; the sequence of the non-coding region of the 5' end linker and the sequence of the non-coding region of the 3' end linker can be transcribed into a cleavage site of a circRNA precursor, and a circRNA is formed through a sequence complementary affinity effect. According to the technical scheme of the application, the circRNA can be continuously produced in the cell-free protein expression, the degradation of the DNA fragment is solved, the protein expression efficiency is improved, and the template can be repeatedly used after the hydrogel is covalently bonded with the template, so that the production cost is further reduced.

This invention relates to the field of database structure and information retrieval technology, specifically to a method and system for dynamic updating of a geographic information (GIS) database. The method includes the following steps: acquiring remote sensing images to extract boundary nodes; calculating multi-temporal location differences to form a change sequence; removing duplicate moving nodes to obtain continuous change information; extracting abrupt boundary changes based on thresholds; and assigning status identifiers to construct a dynamic GIS update structure. In this invention, a boundary structure is constructed based on node location and time identifiers to achieve continuous expression of boundary changes. Spatial differences are used to generate change sequences, improving the accuracy of dynamic boundary identification. By eliminating duplicate moving trajectories, interference information is reduced, enhancing the accuracy of change extraction. Abrupt change identification and boundary reconstruction mechanism is introduced to strengthen the ability to capture abnormal disturbances. Status identifiers are used to map boundary segments, improving the association relationship of boundary attributes and enhancing the expression efficiency and system responsiveness of data updates.

The present application relates to the field of bioengineering, and particularly relates to a mutant of mu-conotoxin and a preparation method and an enzyme composition thereof.The present application provides a mutant of mu-conotoxin, which has an amino acid sequence as shown in SEQ ID NO:1, or an amino acid sequence obtained by substituting, deleting or adding one or more amino acids in the amino acid sequence, and an amino acid sequence which is functionally identical or similar to the amino acid sequence, or an amino acid sequence which is at least 80% identical to the amino acid sequence.The technical route and process provided by the present application greatly improve the production efficiency and quality of mu-conotoxin, are suitable for industrial large-scale production, and have a wide application prospect.

This invention relates to the fields of molecular biology and biotechnology, and provides a reporter vector for visualizing small RNA-target gene regulation. Using a pCOX2 vector as a backbone, it carries a CaMV 35S promoter, a RUBY reporter gene, and the target site sequence of a small RNA target gene, forming a cis-35S-RUBY-target site expression cassette. The CaMV 35S promoter is effectively linked to and drives the expression of the RUBY reporter gene, and the target site sequence is located downstream of the RUBY reporter gene sequence. This invention also provides a method for constructing and applying the aforementioned reporter vector. By constructing a dedicated reporter vector carrying the RUBY reporter gene and combining it with the target site sequence of a small RNA target gene, this invention achieves visual verification of the regulatory effects between small RNA and target genes, offering advantages such as ease of operation, intuitiveness, high efficiency, non-invasiveness, and low cost.

The invention relates to a maltose-based bacillus subtilis self-induced gene expression system and application thereof, and belongs to the field of gene engineering. An induction system commonly used in bacillus subtilis has the problems that exogenous addition is needed, operation is tedious, cost is high, expression intensity is low and the like, and efficient expression and industrialization of protein and metabolites are not facilitated. According to the invention, site-directed mutagenesis is carried out on a PmalA promoter to obtain a high-performance promoter PmalA1, and a T7 RNA polymerase amplification module is further combined to construct a PT7malA1 expression system. The system can automatically start expression after glucose is used up without adding an inducer, and has the advantages of simple and convenient induction process, low leakage, high expression intensity, capability of accurately controlling the induction time by regulating the glucose concentration and the like. By applying the system, efficient synthesis of lycopene and beta-carotene is successfully realized, the yield is remarkably increased, and excellent industrial application potential is shown.

The present application belongs to the technical field of molecular biology, and relates to a truncated fish peptidoglycan recognition protein and application thereof. The amino acid sequence of the truncated fish peptidoglycan recognition protein is shown in Table SEQ ID NO. 1. The truncated fish peptidoglycan recognition protein provided by the present application has higher expression efficiency and higher yield in a prokaryotic expression system compared with a natural peptidoglycan recognition protein. The truncated fish peptidoglycan recognition protein has stronger binding capacity with peptidoglycan (PGN) and lipopolysaccharide (LPS), which are main components of cell walls of gram-positive bacteria and gram-negative bacteria. The bacteriostatic and agglutination capacity of the truncated fish peptidoglycan recognition protein on gram-positive bacteria and gram-negative bacteria is significantly improved. These characteristics show that the truncated fish peptidoglycan recognition protein has development potential as an aquatic immunopotentiator, and provides important support for subsequent application research.

The invention relates to a quantum graph neural network training method, a related method and a related device. The training method comprises the following steps: acquiring a constructed quantum graph neural network model; inputting all nodes in each piece of training data into a quantum feature extractor module to obtain semantic features and attribute features of each node after optimization; performing weighted aggregation on the optimized semantic features of the nodes with the connection relationship to obtain current semantic features of the nodes; according to the original attribute characteristics of the nodes, the optimized attribute characteristics of the nodes are constrained, and the current attribute characteristics of the nodes are obtained; inputting the current semantic features and the current attribute features of all nodes of the training data into a quantum processor network to obtain an output result; and calculating a loss value between the output result and the corresponding training label based on the loss function, updating trainable parameters in the quantum graph neural network model according to the loss value, and carrying out iterative training until the model is converged to obtain a trained quantum graph neural network model.