25 results about "Codon optimization" patented technology

This invention belongs to the field of genetic engineering technology, specifically relating to an engineered bacterium that efficiently synthesizes curcumin using ferulic acid as a precursor, its construction method, and its application. This invention enhances ferulic acid uptake and precursor supply capabilities from the source by knocking out the fdc and mcrC genes and overexpressing the tnaT and accD genes. Furthermore, it achieves high-level stable expression of exogenous key enzyme genes DCS and CURS through codon optimization. Combined with the synergistic effect of a two-stage fermentation regulation process, a stepwise increase in ferulic acid conversion rate is achieved. In a fermentation system with an initial ferulic acid concentration of 5 g / L, the conversion rate of the recombinant strain of this invention increased from 28.6% to 55.2% compared to the unoptimized strain, and further reached 80.2% after fermentation process optimization. Simultaneously, it effectively inhibits the formation of the byproduct 4-vinylguaiacol, significantly improving the utilization rate of ferulic acid and reducing the raw material cost of curcumin biosynthesis.

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.

Provided is a technique relating to codon optimization. A method for optimizing a nucleic acid sequence for expression of a protein in a host comprises: obtaining a protein subsequence-nucleic acid subsequence pair according to collected highly expressed protein sequences and encoding nucleic acid sequences thereof, so as to form a training set; using the training set to train a neural machine translation model, wherein the neural machine translation model is used to realize translation from an amino acid sequence to a codon sequence; cleaving the protein sequence requiring codon optimization into protein subsequences; using the trained neural machine translation model to translate the protein subsequences from amino acid sequences to codon sequences; and overlapping the translated subsequences, so as to combine same into a full-length codon sequence, wherein during overlapping to synthesize the codon sequence, the synonymous codon with the highest occurrence frequency or number is selected as the optimal codon for the position according to the frequency or the number of the synonymous codon corresponding to each amino acid position of the protein sequence.

This invention discloses the application of arginine-depleted engineered bacteria for the treatment of soft tissue sarcomas, including preliminary validation, obtaining qualified engineered bacteria SGR1, obtaining drug compositions and regimens, experimental conclusions, and clinical application promotion. During use, clinical soft tissue sarcoma samples are collected, including undifferentiated pleomorphic sarcoma, synovial sarcoma, and liposarcoma. Immunohistochemistry (IHC) is used to detect ASS1 protein expression, qPCR is used to detect ASS1 gene transcription levels, and the proportion of ASS1 deletion / low expression is statistically analyzed. Arginine auxotrophic sarcoma subpopulations are confirmed based on literature. Cells are cultured in arginine-containing or arginine-free media, and cell viability, colony formation, cell cycle, and apoptosis are detected. The sensitivity of ASS1-deficient cells to arginine deprivation is verified, and therapeutic targets are identified. The enzyme activity, substrate specificity, and immunogenicity of human ARG I / II, mycoplasma ADI, and Pseudomonas ADI are compared. High-activity, low-immunogenic enzymes or enzyme combinations (such as human ARG I + microbial ADI) are selected, and site-directed mutagenesis or codon optimization is performed on microbial ADI to increase expression levels.

The application discloses a recombinant bacterium and a use thereof, the recombinant bacterium is a yeast containing or expressing aai-na-xiang monoterpene synthase BbTPS3 or an ai-na-xiang monoterpene synthase BbTPS3 fusion protein in vivo, and is used for producing levorotatory borneol; experiments prove that the recombinant bacterium after modification (such as codon optimization, protein truncation, Kozak increase or protein fusion) can improve the yield of levorotatory borneol, and is suitable for industrialized production of levorotatory borneol.

This invention belongs to the fields of genetic engineering and biomedicine, specifically relating to a method for codon optimization of human coagulation factor IX and the recombinant human coagulation factor IX encoding gene and its applications. Based on RiboDecode, this invention optimizes relevant parameters to improve the codons of the target gene, thereby increasing the expression level and activity of the target gene in the host. The optimized FIX gene mRNA molecule exhibits good stability and high translation efficiency; further addition of GCCACC upstream of the start codon further enhances expression efficiency. This invention optimizes transfection-related operations, identifies optimal transfection conditions, and achieves maximum expression levels through transient transfection, effectively improving the yield of recombinant human coagulation factor IX. The results of the examples show that the yield of recombinant human coagulation factor IX can reach 1.58±0.10 mg / L, and the coagulation activity can reach 66.8±0.51%, with significant optimization of all key indicators.

This invention belongs to the field of genetic engineering enzyme preparation technology, and particularly relates to a high-expression xylanase, gene editing, recombinant engineered bacteria, and their applications. This invention involves targeted amino acid site mutations in the wild-type xylanase Xyn-WT to obtain five mutant xylanases. Compared to the wild-type xylanase, the enzyme activities of all five mutants are significantly increased. Among them, the Xyn-4 mutant exhibits the best enzyme activity, reaching 245.17 U / mL, an increase of 90.60% compared to the wild type. Codon optimization further increased the expression level by 20.74%, demonstrating both high expression and excellent catalytic activity. The xylanase of this invention has high yield and high enzyme activity, making it suitable for industrial production scenarios such as feed, food, pulp and paper making, biomass degradation, and textiles, and possessing extremely high industrial application value.

This invention belongs to the fields of genetic engineering and enzyme engineering, specifically relating to a chitinase mutant ChiTgM and its applications. This invention obtains the chitinase ChiTg through gene cloning, achieves heterologous expression of it in Pichia pastoris through codon optimization, and obtains the chitinase mutant ChiTgM through site-directed mutagenesis and combinatorial mutagenesis. Analysis of the enzymatic characteristics of chitinase ChiTg and the mutant ChiTgM shows that, compared to the starting template ChiTg, the chitinase mutant ChiTgM provided by this invention exhibits a 1.81-fold increase in specific activity and demonstrates excellent hydrolytic properties. Its application in the enzymatic preparation of chitin oligosaccharides can efficiently hydrolyze colloidal chitin, laying the foundation for its further industrial application.

The application discloses a Marburg virus-based micro-genome replication-defective system and application thereof, and belongs to the field of biological medicine. The system significantly improves the expression efficiency and signal-to-noise ratio of a model reporter gene through a codon optimization strategy. Experimental verification shows that the model can safely simulate the MARV genome replication and transcription process under a biosafety level 2 (BSL-2) condition, and has high specificity and sensitivity. Through parallel verification of positive drugs such as ribavirin, suramin sodium salt and negative controls such as oseltamivir, it is confirmed that the model can accurately and quantitatively evaluate the activity of small-molecule drugs targeting the replication link of the virus and calculate EC50. The model provides a safe and efficient platform for the study of the MARV replication mechanism and the high-throughput screening of antiviral drugs, breaks through the limitation of a biosafety level 4 laboratory, and has important scientific research and application value.

This invention discloses a recombinant yeast for producing α-ionone, its construction method, and its applications. The construction method involves using a yeast encoding osmanthus flowers... Osmanthus fragrans The gene for the carotenoid cleavage dioxygenase OfCCD1 was obtained through rational modification and codon optimization. OfCCD1m The gene, with its 5' end fused to the N-terminal membrane-anchored peptide coding sequence of human tyrosine kinase LCK, was transformed into the δ site of Saccharomyces cerevisiae TW3 to obtain recombinant yeast TW4 producing α-ionone. The Saccharomyces cerevisiae strain TW4 producing α-ionone of this invention has the following beneficial effects: it achieves de novo synthesis of α-ionone in Saccharomyces cerevisiae for the first time, filling a technological gap in this field; by using a membrane anchoring strategy to locate the OfCCD1m enzyme in the yeast plasma membrane, the contact efficiency between the enzyme and the substrate ε-carotene is significantly improved, resulting in an α-ionone yield of 1.46 mg / L.

The application provides an RNA, an RNA combination and application thereof, and a multivalent monkeypox vaccine, and relates to the technical field of biotechnology.The mRNA provided by the application is obtained after codon optimization, and can express antigen proteins in vitro cells with high efficiency; the mRNA combination provided by the application has high expression, and after immunizing mice, high-titer neutralizing antibodies against the live virus of the vaccinia virus Tianlu strain can be detected, and the mRNA combination has good immunogenicity, and is of great significance for the prevention of the monkeypox virus.The multivalent monkeypox mRNA vaccine provided by the application can realize stable and safe expression in vivo and effectively activate an immune response, and can cause a neutralizing antibody response, and in the serum of the immunized mice, high-titer neutralizing antibodies against the live virus of the vaccinia virus Tianlu strain can be detected.

This invention provides an optimized method for enhancing self-amplifying RNA expression and an enhanced self-amplifying RNA. Specifically, it provides an optimized method for enhancing self-amplifying RNA expression, which involves codon optimization. The optimized region is a specific region of the alphavirus non-structural protein sequence of the self-amplifying RNA, where the alphavirus non-structural protein is nsP1234. It also provides a method for preparing the enhanced self-amplifying RNA, an enhanced self-amplifying RNA, and a plasmid. This application improves the expression efficiency of the self-amplifying RNA target gene by designing and constructing codon sequence optimization for certain regions of these non-structural proteins.

The application discloses application of asd gene and mutants thereof in preparation of lysine, and belongs to the technical field of genetic engineering. The technical problem to be solved by the application is how to improve the fermentation yield of lysine, for example, the yield of lysine catalyzed by whole cells of microorganisms. In order to solve the technical problem, the application provides application of aspartate semialdehyde dehydrogenase in improving the lysine yield of microorganisms. The application also provides a recombinant microorganism containing aspartate semialdehyde dehydrogenase and provides a method for preparing lysine by using the recombinant microorganism. The application first discloses the use of aspartate semialdehyde dehydrogenase and the coding gene thereof in improving the L-lysine yield of microorganisms. The application obtains an optimized coding gene of aspartate semialdehyde dehydrogenase by codon optimization and produces unexpected technical effects, and has great application potential in the field of microbial fermentation of lysine.

The invention discloses a codon optimization method and device.According to the codon optimization method, after an initial codon sequence corresponding to an input protein sequence is obtained, a naturalness score and a protein high expression probability are jointly used as optimization targets, iterative optimization is conducted on the initial codon sequence, and then the initial codon sequence is obtained; and obtaining a target codon sequence matched with the protein sequence. According to the method, the naturalness score and the high expression probability of the codon are jointly used as optimization targets, iterative optimization is carried out on the codon sequence, and the codon sequence which not only conforms to the context compatibility of the host codon but also has the high expression characteristic can be obtained.

This invention relates to the field of vaccine technology and provides a CHO cell line for the TGEV strain S protein and its application. The CHO cell line has a TGEV S protein encoding gene with codon optimization carrying the TPA signal peptide integrated into its chromosome. The nucleotide sequence of the TGEV S protein encoding gene is shown in SEQ ID NO.1. This invention not only significantly improves the exocrine expression efficiency and stability of the TGEV S protein but also increases protein quantity and yield, establishing an expression system that can operate stably for a long time and is suitable for large-scale production, demonstrating clear technological advancement and practical application value.

The present invention relates to a method for producing a microorganism having N-deacetylation activity and N-sulfation activity, comprising the steps of (I) modifying the nucleotide sequence of DNA encoding N-deacetylase / N-sulfotransferase and (II) introducing the DNA containing the modified nucleotide sequence in step (I) into a microorganism so that it can express, wherein step (I) is a step of codon optimization to match the codon usage frequency of a different species of organism than the microorganism. The present invention also relates to a method for producing a microorganism having N-deacetylation activity and N-sulfation activity, comprising the step of (i) expressing a specific protein in the microorganism.

The present invention provides compositions and methods for making circular RNAs, in particular circular RNAs encoding one or more proteins and having superior stability. Such circular RNAs comprise complementary regions that are capable of forming double-stranded regions that are important for maintaining the stability of the circular RNA under different environments and that facilitate translation and circularization. Such complementary regions can be formed by codon optimization or can be introduced as heterologous sequences. The present invention also provides circular RNAs that express self-amplifying RNA non-structural proteins, e.g., (nsP1-4), which can amplify independent mRNA molecules containing corresponding conserved sequence elements (CSEs). These mRNAs can encode a gene of interest (GOI). When they are introduced together into a target cell, the circular RNAs can amplify the mRNAs, thereby enabling stable high expression of the gene of interest.

The application belongs to the technical field of biotechnology, discloses mRNA and a preparation method, use and vaccine thereof, a nucleotide sequence of a HA antigen coding gene of the mRNA is shown as SEQ ID NO:1, the mRNA is obtained by the following steps: adding a T7 promoter, a 5' end untranslated region and a Kozak sequence at the front end of a coding sequence of an HA protein of a swine influenza virus H1N2 after codon optimization, adding a 3' end untranslated region at the rear end, forming a DNA template sequence, and then obtaining the mRNA, and the vaccine prepared by using the mRNA has the advantages of good immunogenicity and strong protection.

The application discloses a chikungunya fever nucleic acid vaccine and belongs to the technical field of nucleic acid vaccines. A transmembrane protein sequence is obtained through multiple recombination optimizations, the protein sequence is connected with E2 and E1 protein sequences, a polyprotein is constructed, and the E2 and E1 proteins of chikungunya fever can be efficiently expressed through codon optimization and circular mRNA or linear mRNA technology. Compared with original virus protein sequences, the expression efficiency is improved by 2.7 times, the antibody titer of a mouse immunized by the circular mRNA vaccine is improved by 4.7 times, and the antibody duration is longer.

The present application relates to the field of plant genetic engineering technology, and particularly relates to the application of algal NySAT2 gene in regulating alfalfa crude protein and washing fiber. The present application uses gene synthesis and codon optimization technology to obtain the sequence of the coding region of the codon-optimized NySAT2 gene, constructs an algal NySAT2 gene overexpression vector, introduces the target gene into the alfalfa genome through the Agrobacterium rhizogenes-mediated genetic transformation method, and obtains the transgenic plant using the genetic transformation system. Through molecular identification and phenotype analysis, it is found that the alfalfa plants overexpressing the NySAT2 gene have a significantly increased crude protein content compared with the wild-type alfalfa plants, while the contents of acid detergent fiber and neutral detergent fiber are significantly reduced, effectively improving the forage quality and feeding value of alfalfa.

This invention discloses a chitin deacetylase product and its application in agricultural disease prevention. Through codon optimization, the *Rhodococcus* chitin deacetylase gene (SEQ ID NO:1) was cloned into the *Pichia pastoris* expression vector pGAPZa A, constructing the recombinant vector pGAPZaA-CDA-His, which was then transformed into the *Pichia pastoris* SMD1168 host, achieving efficient secretory expression of the enzyme. The resulting recombinant enzyme exhibits high activity and good thermostability. This invention includes a universal enzymatic deacetylation process characterized by deacetylation efficiency, applicable to chitin deacetylases from various sources. This invention also provides a method for further enzymatic hydrolysis to produce N-acetylglucosamine monomers or their oligosaccharides, and the application of the recombinant enzyme and its product in the preparation of biopesticides and the treatment of chitin-containing biomass materials.

The application discloses a kind of for synthesizing β - terpene synthase CcTPS6 of eucalyptol and its encoding gene and its product β - the application of eucalyptol as anti-aging drug belongs to synthetic biology and natural medicine chemistry technical field. The application starts from Labiatae plant torch flower ( Colquhounia coccinea var. mollis ) Cloning and functional identification of a synthetic β - eucalyptol single-function terpene synthase CcTPS6 encoding gene, its nucleotide sequence is as shown in Seq ID No.2, after codon optimization and site-directed mutagenesis, connect with expression vector pCold-TF, construct into the recombinant plasmid capable of expressing in escherichia coli, again the recombinant plasmid is transformed into escherichia coli and is constructed into engineering cell, realizes the heterologous high-efficiency synthesis compound β - eucalyptol in escherichia coli. The gene engineering cell constructed by the application is safe and stable, and the production cycle is short, which shows its great value in application development. The CcTPS6 product provided by the application β - eucalyptol has anti-aging activity and can be applied in the preparation of anti-aging drugs.

This invention relates to a Pichia pastoris chassis for efficiently supplying the terpene precursor IPP and its construction method, belonging to the field of molecular biology. The invention involves screening three highly expressed HMGR genes from the latex tissue of the Brazilian rubber tree using transcriptome sequencing. After codon optimization in Pichia pastoris, these genes were assembled into an expression cassette with a screening marker, a bleomycin resistance gene, expression elements, and a weak promoter. Using two fragments from different upstream regions of the CDS of the endogenous EGR9 gene in Pichia pastoris as homologous arms, the expression cassette was integrated into the Pichia pastoris genome through homologous recombination, replacing the original ERG9 gene promoter to weaken the competitive consumption of IPP by the squalene synthesis pathway. The constructed Pichia pastoris mutant strain showed a 430% higher IPP content than the wild type, with no significant difference in growth rate. It efficiently supplies IPP, providing a sufficient precursor for the heterologous synthesis of terpenoids, and has significant industrial application value.

The application provides a TaMn-SOD protein and heterologous expression and application thereof, and belongs to the technical field of oxidoreductases. The TaMn-SOD protein comprises an amino acid sequence shown in SEQ ID NO:1, is based on the full sequence amino acid of TaMn-SOD of aquifex pyrophilus, and is obtained through codon optimization, heterologous expression, extraction, purification, dialysis and concentration. The TaMn-SOD protein has multiple characteristics such as high-temperature resistance, acid and alkali resistance and the like.

The present disclosure provides a highly stable and recombinant humanized elastin, a genetically engineered bacterium expressing the elastin, and an application therefor, and belongs to the technical field of connective tissue peptides. A nucleotide sequence of the recombinant humanized elastin is as shown in SEQ ID NO.1; and an amino acid sequence is as shown in SEQ ID NO.4. The present disclosure performs codon optimization on the nucleotide sequence of the humanized elastin and then transfer the nucleotide sequence into Escherichia coli to construct a genetically engineered bacterium expressing the recombinant humanized elastin, which can increase an expression level of the recombinant humanized elastin, improve stability of the recombinant humanized elastin, increase a migration rate of the recombinant humanized elastin to HSF cells, and increase gene expression levels of h-ELN, h-COL1A1, and h-COL3A1 in HSF cells. 20 25 20 37 68 21 M ay 2 02 5 A B S T R A C T 2 0 2 5 2 0 3 7 6 8 2 1 M a y 2 0 2 5 1 / 2 FIG. 1 FIG. 2 1 / 2 M 1 2 3 4 kDa 180 130 100 70 55 40 35 25 15 FIG. 1 Oh 24h HSF Oh 24h NC rhELN-optimization 1 rhELN-optimization 2 rhELN-pre-optimization FIG. 2 20 25 20 37 68 21 M ay 2 02 5 k D a 1 8 0 2 0 2 5 2 0 3 7 6 8 2 1 M a y 2 0 2 5 5 5 3 5 2 5 F I G . 1 O h 2 4 h H S F O h 2 4 h N C