49results about How to "Improve expression efficiency" patented technology

The invention discloses a lightweight bearing surface defect detection method and system based on improved YOLOv10n. The method comprises the following steps: acquiring a bearing surface defect sample data set; the method comprises the following steps: improving a YOLOv10 network structure to obtain an FM-NET algorithm model; training an FM-NET algorithm model based on the bearing surface defect sample data set to obtain a target FM-NET algorithm model; and performing defect detection on a to-be-detected bearing image according to the target FM-NET algorithm model, and determining a defect position and a corresponding defect category in the bearing image. According to the method, the multi-scale feature extraction capability and the model expression efficiency are remarkably improved, the robustness of the model in a complex scene is enhanced, the reasoning speed is remarkably improved on the premise of keeping high precision, and therefore good balance between the precision and the efficiency is achieved.

The invention relates to the technical field of bioinformatics, in particular to a mining method and system for synthetic biology functional elements and a storage medium. The method comprises the following steps: acquiring functional demand data of a target metabolic pathway; performing evolutionary distance analysis and function prediction on homologous genes of a plurality of species according to the function demand data, and performing protein structural domain conservative classification on preset genome sequence data to obtain candidate element cluster data with function relevance; obtaining expression system data of the host cells; and according to the candidate element cluster data and the expression system data, scoring each functional element based on codon optimization degree, protein folding stability and metabolic load prediction to obtain hierarchical element suitability data. According to the method, factors such as sequence characteristics, expression efficiency and regulation and control network interaction are considered at the same time, multi-level accurate screening is achieved, and therefore the low efficiency and limitation of a traditional screening mode are overcome.

The invention relates to the technical field of microorganisms, in particular to recombinant pichia pastoris for expressing plastic degrading enzyme LCCICCG as well as a construction method and application of the recombinant pichia pastoris. According to the invention, pichia pastoris is taken as a host cell, the coding gene lccICCG of the plastic degrading enzyme is heterologously expressed, and single-copy or multi-copy recombinant pichia pastoris is constructed. By optimizing culture conditions, the expression quantity and biomass of the plastic degrading enzyme protein are remarkably improved; by optimizing a fermentation strategy, high-density fermentation production of the LCCICCG protein in a 5L fermentation tank is realized, and a technical basis is provided for realizing large-scale industrial production of the protein. The improvements not only improve the production efficiency and reduce the cost of industrial production, but also provide powerful support for commercial application of the plastic depolymerases.

The invention discloses a deep mutation scanning method based on protein-protein interaction in bacterial cells and a binding affinity prediction method. In the invention, split dCas9 proteins (SEQ ID NO.1 and SEQ ID NO.2) are respectively subjected to fusion expression with a receptor and a ligand with binding affinity to be detected, and are combined with a toxic sgRNA guide sequence (SEQ ID NO.3), and when PPI occurs, a toxic dCas9: sgRNA complex is formed to inhibit bacterial growth, so that a PPI affinity size signal is converted into a bacterial cell generation speed signal, and the signal can be quantified into a copy number value through next-generation sequencing. The invention further designs a deep learning binding affinity prediction model, high-affinity sequences can be screened from theoretical mutation space, the workload and cost of experimental verification are remarkably reduced, and a reliable basis is provided for subsequent protein function optimization and design.

The invention discloses an rhFN recombinant human chemical fibronectin process of a genetic engineering technology, and particularly relates to the technical field of genetic engineering, and the rhFN recombinant human chemical fibronectin process comprises target gene acquisition, gene cloning and vector construction, host cell selection and transformation, cell culture and screening, protein expression and optimization, and protein separation and purification. According to the rhFN recombinant human chemical fibronectin process of the genetic engineering technology provided by the invention, high-quality target genes are accurately obtained through technologies such as new-generation sequencing and gene synthesis, and the gene expression efficiency and stability are enhanced by virtue of optimized vector construction and regulatory elements; protein expression ability and specificity are improved in host cell selection and transformation, efficient culture and accurate screening are realized in cell culture and screening links, total yield is increased in a protein expression optimization link, purity and efficiency are improved in a protein separation and purification link, and product quality and safety are ensured through comprehensive quality control. The requirements in the fields of medical treatment and scientific research are expected to be better met.

The present invention relates to a vaccine framework fusion protein, a recombinant vaccine and its application. The fusion protein of the present invention comprises the intracellular segment of the low-affinity immunoglobulin gamma Fc region receptor II (FcγRII) and the WW domain (WW domain) of the E3 ubiquitin protein ligase Nedd4 family member protein, and / or one or more antigens, and / or one or more transmembrane regions. The present invention provides a new recombinant vaccine molecular architecture, which has a better ability to promote the expression and secretion of immune antigens and improve the effect of immune level, provides new ideas and methods for the design of recombinant vaccines, and can be widely used in the field of vaccines.

The invention belongs to the technical field of biology, and particularly relates to recombinant III-type humanized collagen as well as a preparation method and application thereof. On the basis of the III-type human collagen amino acid sequence, the amino acid sequence is designed by means of bioinformatics and the like, so that the recombinant collagen realizes higher expression efficiency and a triple helix structure on the premise of maintaining high biological activity, a section of brand new collagen sequence containing 180 amino acids is designed, the protein molecular weight is small, and the recombinant collagen has the advantages of high expression efficiency, high yield, high yield and the like. The skin permeability is better. Compared with a commercially available collagen product, the recombinant humanized collagen prepared by the technical scheme has better stability, cell proliferation and migration promotion effect and barrier repair capability, so that the recombinant humanized collagen has good practical application value.

The invention discloses a yak intramuscular precursor fat immortalized cell line and a preparation method thereof. The method comprises the following steps: (1) obtaining yak primary intramuscular precursor fat cells; (2) performing SV40T lentivirus infection on the precursor fat cells in the yak muscle and screening cell strains: performing SV40T lentivirus infection by utilizing a 1 / 2 small-volume infection method to obtain immortalized cells after the precursor fat cells in the yak primary muscle are infected with SV40T, namely immortalized cells for short; puromycin is adopted to screen immortalized cells; and diluting the screened cells by using a limited dilution method, and carrying out enlarged culture to obtain the immortalized cell line. The yak intramuscular precursor fat immortalized cell line constructed by the invention has genetic homogeneity, the heterogeneity influence of cell populations is eliminated, and an in-vitro cell model is provided for subsequent research related to yak intramuscular fat cells.

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 present invention belongs to the field of genetic engineering technology, and discloses a pSFV-p54 replicon particle and a preparation method and application thereof. The preparation method of the pSFV-p54 replicon particle comprises the following steps: (1) using pSFV-sp6-EGFP plasmid and flag-p54 gene as materials, constructing a pSFV-sp6-p54 plasmid; the flag-p54 gene is as shown in SEQ ID NO: 1; (2) linearizing the SFV-helper auxiliary plasmid and the pSFV-sp6-p54 plasmid respectively and performing in vitro transcription, and co-electrotransferring the obtained mRNA into BHK cells or Vero cells to obtain pSFV-p54 replicon particles. The present invention uses Semliki Forest virus as a vector, inserts the African swine fever p54 gene, and the prepared pSFV-p54 replicon particles can efficiently express the African swine fever virus p54 protein, and have high biosafety, and can be used to develop African swine fever related vaccines.

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 provides a method for synthesizing a cobra antibacterial peptide NA30-CATH through multi-copy integration and high-density fermentation and application of the cobra antibacterial peptide NA30-CATH, and belongs to the technical field of genetic engineering and bioengineering. According to the invention, a series-connection two-copy plasmid is constructed, pichia pastoris is taken as a host, two-copy series-connection protein is successfully expressed, and the expression yield of the cobra antibacterial peptide NA30-CATH is increased by more than two times. According to the present invention, the cobra antibacterial peptide NA30-CATH can be synthesized at a high level by using the cobra antibacterial peptide NA30-CATH and constructing and optimizing the two-copy integration method and optimizing the fermentation process, and the method has advantages of strong versatility, high expression efficiency, biological activity, low cost, easy large-scale production, and the like. In addition, the invention further provides application of the cobra antibacterial peptide NA30-CATH as an antibacterial agent in the process of breeding the largemouth bass, and the survival rate of the largemouth bass is increased.

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 application belongs to the field of plant genetic engineering, and particularly relates to a series of expression vectors designed based on Capsicum annuum vein yellowing virus (PoPeVYV). The application proves that co-transfection of PoPeVYV aRNA and the expression vector can improve the expression efficiency of exogenous proteins in an infected object. The vector can not only efficiently express exogenous proteins in a host, but also realize assembly of virus-like particles. In summary, the application develops a new virus vector and a co-expression system thereof, and provides a new selection for a plant expression system.

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 present application relates to an ATP-responsive lipid, a nucleic acid nano-preparation and a preparation method. The structural formula of the ATP-responsive lipid is shown in the following formula (I). The ATP-responsive lipid and the nucleic acid nano-preparation of the present application can be combined with ATP under the condition of high ATP level in cells, increase the fluidity of the membrane, promote the fusion of the lipid nanoparticle and the endosome membrane, destroy the integrity of the endosome membrane, increase the efficiency of lysosome escape, and then improve the efficiency of nucleic acid delivery and expression.

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 present application relates to a kind of based on topological-frequency domain collaborative field sensing network's remote sensing image semantic analysis method, belong to remote sensing image semantic analysis technical field.The method includes: remote sensing image is initialized pretreatment, and image field representation is obtained;Topological-frequency domain collaborative field sensing encoder is constructed, and image field representation is carried out multi-level field decomposition and reorganization, and deep field representation is obtained;Field dynamic aggregation bottleneck layer is constructed, and deep field representation is carried out global field perception and sparse screening, and global optimization field feature is obtained;Field cognitive decoder is constructed, and global optimization field feature is inverted into pixel-level semantic field;Field cognitive loss function is constructed, and pixel-level semantic field is combined by back propagation training topological-frequency domain collaborative field sensing network, and the semantic field graph is output using trained topological-frequency domain collaborative field sensing network.It aims at solving the technical problems that existing technology has single cognitive dimension, feature fusion mean value, structure integrity loss and weak generalization ability.

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a triglyceride-derived ionizable lipid compound, a preparation method thereof, a drug delivery carrier and application. The chemical structural formula of the ionizable lipid compound derived from triglyceride is as shown in a formula (I): # imgabs0 # formula (I). The ionizable lipid compound has the advantages of being high in degradability, high in safety, beneficial to release of carried nucleic acid drugs and the like, and a drug delivery carrier containing the compound has the excellent performance of being good in in-vivo targeting, high in biological safety, high in mRNA expression efficiency and the like. Targeted organs and tissues of the drug delivery carrier containing the compound comprise tumors, heart, liver, spleen, lung, kidney and the like.

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.

The invention relates to a construction method of an MN1-BEND2 fusion gene knock-in mouse animal model and application of the MN1-BEND2 fusion gene knock-in mouse animal model as an astroblastoma animal model. According to the method disclosed by the invention, the MN1-BEND2 fusion gene knock-in mouse animal model can be obtained at a relatively high success rate, the expression of the MN1-BEND2 fusion gene in the obtained mouse animal model is cell-specific, and the MN1-BEND2 fusion gene positive cells have the proliferation state and characteristic molecular phenotype of the astroblastoma, so that the MN1-BEND2 fusion gene knock-in mouse animal model has the advantages that the MN1-BEND2 fusion gene knock-in mouse animal model has a good application prospect. The expression of the gene in a specific area can well simulate the astroblastoma, so that an important research tool is provided for mechanism research and drug development of the astroblastoma.

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.

The invention relates to the technical field of biology, in particular to EGFR polypeptide and application thereof. The novel truncated EGFR polypeptide is composed of extracellular structural domains III and IV, a transmembrane region and an intracellular proximal membrane terminal amino acid sequence, does not contain an extracellular ligand binding structural domain and an intracellular tyrosine kinase structural domain, and is named as JxM-EGFRt. Compared with the traditional EGFRt, the JxM-EGFRt improves the expression efficiency and the expression abundance of the EGFR polypeptide while improving the safety, on one hand, the ADCC efficiency is enhanced, the safety is better, and on the other hand, the truncated EGFR polypeptide can be used as a selection marker, and the marking efficiency is improved. Besides, CAR-T cells expressing the JxM-EGFRt polypeptide have a higher initial / immature T cell proportion, in-vivo and in-vitro storage of target cells is enhanced, and the in-vivo and in-vitro anti-tumor effect is improved.

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 invention discloses a histone gene-derived bidirectional promoter for Japanese oryzias latipe and application of the histone gene-derived bidirectional promoter. The bidirectional promoter provided by the invention can simultaneously drive the expression of genes on two sides, the efficiency of a multi-gene expression system is effectively improved, the fragment is only 353 bp, is short, small and simple, is convenient to introduce in different vectors and expression systems, reduces the occupation of the vector capacity, and is higher in flexibility. The gene is derived from a histone intergene region, has relatively strong transcriptional activity, can maintain high-level expression in various cell types, and has relatively strong application advantages in gene expression in the fields of aquaculture, ecological research, fish biology and the like.

The invention provides a preparation method of a two-copy perch antibacterial peptide wb-moronecidin, and belongs to the technical field of genetic engineering and bioengineering. According to the invention, the plasmid with two tandem copies is constructed, pichia pastoris is taken as a host, the two-copy tandem protein is successfully expressed, and the expression yield of the wb-moronecidin of the perch antibacterial peptide is improved. According to the present invention, the perch antibacterial peptide wb-moronecidin can be synthesized at a high level by using the two-copy integration method and the fermentation process optimization, and the method has advantages of strong versatility, high expression efficiency, biological activity, low cost, easy large-scale production, and the like, and can be used for the preparation of the perch antibacterial peptide wb-moronecidin in the perch antibacterial peptide wb-moronecidin in the perch antibacterial peptide wb-moronecidin.

The present invention provides a multispecific antigen binding protein comprising at least: (a) a first binding domain that specifically binds to a natural killer cell activated receptor (NKp46); and (b) a second binding domain that specifically binds to mesothelin (MSLN). The multispecific antigen binding protein provided by the invention can be bound with NKp46 and MSLN with high affinity, so that immune cells expressing the NKp46 are mediated to kill MSLN positive cells; and the multispecific antigen binding protein can have a full-length IgG configuration, so that the multispecific antigen binding protein has the advantages of a natural antibody in application. The invention also provides application of the multispecific antigen binding protein in tumor treatment.

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.