142 results about "Codon optimized" patented technology

The invention discloses an SaRNA vaccine for echinococcosis as well as a preparation method and application of the SaRNA vaccine. The preparation method of the SaRNA vaccine comprises the following steps: carrying out codon optimization on a modified target antigen protein through a genetic engineering technology, then assembling the modified target antigen protein with a self-replicating protein sequence, 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a plasmid, and carrying out purification to obtain the SaRNA vaccine. The preparation method comprises the following steps: constructing recombinant plasmids, sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmids to prepare SaRNA molecules, and finally wrapping the SaRNA molecules in lipid nanoparticles to form the SaRNA vaccine for the echinococcosis. Experiments prove that the SaRNA vaccine can activate humoral immunity and cellular immunity of mice at the same time, high-level EG95 specific antibodies and cytokines can be generated through low-dose immunity, and the SaRNA vaccine has wide application prospects in the aspect of preventing and / or treating the echinococcosis.

The present disclosure provides codon optimized GBA1 polynucleotides encoding a GCase protein, wherein a portion of the coding sequence deviates from the wild type. The disclosure also provides expression constructs, vectors, viral particles or compositions containing the disclosed polynucleotides. In addition, methods and uses of these polynucleotides, expression constructs, vectors, viral particles or compositions are also provided, including the treatment of diseases or conditions associated with GCase deficiency.

The present invention provides, among other things, improved pharmaceutical compositions comprising codon-optimized mRNA encoding a peptide or polypeptide encapsulated in a lipid nanoparticle comprising one or more of the cationic lipids that are particularly effective for pulmonary delivery.

The invention belongs to the technical field of enzyme engineering, and particularly relates to a codon optimized superoxide dismutase AfSOD gene and application thereof in degradation of aflatoxin B1 and zearalenone. Under the conditions that the pH is 7, the temperature is 40 DEG C and the enzyme addition amount is 10 micrograms, the degradation rate of the AFB1 reaches 99.08% within 24 hours, and the degradation rate is kept to be 80% or above within 40-80 DEG C; under the conditions that the pH is 8, the temperature is 60 DEG C and the enzyme addition amount is 20 micrograms, the degradation rate of ZEN reaches 85.1% in 24 hours, and the degradation rate is kept to be 80% or above within 50-80 DEG C; the result shows that AfSOD is a toxin degrading enzyme which can tolerate a certain degree of high temperature; the degradation rates of AFB1 and ZEN in polluted peanut powder or corn powder are 73.7% and 88.2% respectively; the invention provides a new candidate for detoxification of AFB1 and ZEN in food or feed, and has a good application prospect.

The invention belongs to the technical field of bioengineering, and discloses a high-yield hyaluronic acid bacillus subtilis strain for expressing a hyaluronic acid synthase gene, a construction method and application, and the construction method comprises the following steps: inserting a codon-optimized sphaasA gene into a lytH site in a bacillus subtilis 168 strain in a traceless manner by utilizing a CRISPR / dCas9 system; the preparation method comprises the following steps: synthesizing hyaluronic acid oligosaccharides, simultaneously integrating genes for expressing a synthetic pathway of hyaluronic acid oligosaccharides, reducing the transcriptional level of zwf and pfkA genes, weakening the metabolic flux of a competitive pathway, and finally increasing the expression of sphaasA and tuaD genes according to a ratio of 2: 1 by using plasmids and the interaction of protein scaffolds RIAD and RIDD. When the strain is fermented for 48 hours in a 5L fermentation tank, the yield of the hyaluronic acid reaches 38.02 g / L, and the average molecular weight HA Mw of the low-molecular-weight hyaluronic acid is about (5-13) * 10 < 4 > Da.

The invention discloses a piglet intestinal health targeting probiotic engineering bacterium compound as well as preparation and application thereof. The compound is FP (at) EcN-pnirNKL, wherein a) the EcN-pnirNKL is an engineering strain which is constructed by taking escherichia coli Niss1917 (EcN) as a host and carrying a recombinant plasmid pnirBMisL-NK-Lysin, and a) the EcN-pnirNKL is an engineering strain which is constructed by taking escherichia coli Niss1917 (EcN) as a host and carrying a recombinant plasmid pnirBMisL-NK-Lysin; b) the recombinant plasmid pnirBMisL-NK-Lysin comprises an NK-Lysin gene optimized by a codon, and the NK-Lysin gene is integrated into the anaerobic inducible plasmid pnirBMisL through homologous recombination; c) the FP is a composite nano-particle which takes nano mesoporous silica (MSN) as a core and is formed by crosslinking fucoidin and rhodiola rosea polysaccharide; and d) the FP (at) EcN-pnirNKL is an oral delivery system which is formed by encapsulating the engineering bacterium EcN-pnirNKL in the FP nano particles. When the compound is applied to prevention and treatment of stress response of weaned piglets, various typical symptoms, including weight loss, colon shortening, histopathological injury and colitis disease response, of the piglets induced by dextran sodium sulfate can be remarkably relieved, and meanwhile, the integrity and functions of an intestinal barrier structure are effectively protected.

Disclosed herein is a method for the treatment of Pompe Disease comprising administering a recombinant AAV (rAAV) vector comprising a rAVV genome comprising a heterologous nucleic acid encoding a GAA signal peptide or portion thereof, a heterologous signal peptide, and an acid alpha-glucosidase (GAA) polypeptide, or N-terminal truncation thereof, where the heterologous nucleic acid is operatively linked to a liver-specific promoter, where the nucleic acid encoding GAA polypeptide can be wild type nucleic acid sequence, or modified nucleic acid sequence, or a codon optimized nucleic acid sequence, and can optionally be modified to reduce or completely eliminate CG and CpG dinucleotides and, optionally eliminated alternative reading frames (ARF) content.

The invention discloses a macrophage-targeting lipid nanoparticle and application thereof in malignant tumor treatment, ID3 mRNA is delivered through the lipid nanoparticle, firstly, an ID3 sequence is subjected to codon optimization to improve the expression efficiency and stability of ID3 protein, and the core treatment effect is enhanced; the lipid composition and the targeting ligand of the nanoparticles are further optimized, and mannose modified PEG lipid material (DSPE-PEG2000-Mannose) is specifically combined with the CD206 receptor on the surface of the macrophage, so that the targeting property and the transfection efficiency of the delivery system are remarkably improved. The ID3 mRNA is delivered to the macrophages by utilizing the lipid nanoparticles of the targeted macrophages, so that the tumor cells can be effectively swallowed and killed, the in-situ treatment of pancreatic cancer is realized, and better tumor inhibition and immune activation effects are achieved.

The invention provides a method for synthesizing melanin nanoparticles by using escherichia coli, and belongs to the technical field of biological genetic engineering. According to the method, a bacillus megaterium-derived tyrosinase gene Tyr1 (SEQ ID NO.2) after codon optimization is taken as a target gene, a recombinant expression plasmid pET-24a-Tyr1 is constructed, and the recombinant expression plasmid pET-24a-Tyr1 is transformed into an escherichia coli BL21 (DE3) competent cell to obtain the genetically engineered bacterium. After the engineering bacterium induces expression of tyrosinase through IPTG (isopropyl-beta-d-thiogalactoside), polymerization of an L-tyrosine substrate is catalyzed in the presence of CuSO4 to obtain high-purity melanin nanoparticles. The melanin nano-particles synthesized by the method are uniform in particle size, have wide-spectrum strong absorption characteristics in a wavelength range of 400-800nm, show excellent photo-thermal performance, free radical scavenging capacity and biocompatibility, and have wide application prospects in the fields of biomedicine and functional materials.

The application belongs to the field of biology and relates to a nucleic acid molecule encoding an anti-HER2 antibody, a vector containing the nucleic acid molecule and an expression method, wherein the nucleic acid molecule contains codons optimized for expression cells. By using the nucleic acid molecule of the application, the expression amount of the anti-HER2 antibody in cells can be improved, the yield of the anti-HER2 antibody is increased, and the production cost is reduced; and the anti-HER2 antibody produced has similar performance to that of the original research drug and has a good drug-making prospect.

The present application relates to the field of molecular biology, in particular to a recombinant human heparin N-deacetylase encoding gene, a recombinant vector, a recombinant strain and application thereof, the recombinant human heparin N-deacetylase constructed in the expression process shows excellent stability and activity, has the ability of specific catalysis of deacetylation of acetyl group in heparin precursor, and can effectively generate deacetylated heparin precursor. This property significantly improves the conversion efficiency and reduces the generation of by-products. The present application optimizes the codon of the recombinant human heparin N-deacetylase, and uses the promoter P AOX1 or P GAP to regulate the expression, so that the expression level of the recombinant human heparin N-deacetylase in Pichia pastoris is significantly improved, and the obtained GS115-AOX1-hNDase and GS115-PGAP-hNDase strains successfully realize the secretory expression of the target gene, providing a new strategy and method for the biosynthesis of heparin.

The application discloses a codon-optimized Cas12i3 protein coding gene and application thereof. Specifically disclosed is a DNA molecule, and a nucleotide sequence of the DNA molecule is SEQ ID No. 3. The application also discloses a gene editing system containing the DNA molecule and a method for improving the efficiency of Cas12i3 protein-mediated gene editing. In order to better apply the Cas12i3 protein to mammalian gene editing, different codon optimization schemes are designed, and through software evaluation and experimental verification, a codon optimization scheme with good Cas12i3 expression and editing effect is compared and screened. The application screens the codon optimization scheme for efficiently expressing the Cas12i3 protein in mammals, which can greatly improve the editing efficiency of the Cas12i3 protein, lays a foundation for the wide application of the Cas12i3 protein in mammals, and has great practical application value.

The present disclosure relates to codon-optimized sequences coding for hGCDH polypeptide and recombinant adeno-associated virus (rAAV) vectors comprising one of said sequences under the control of a promoter component. Also provided herein are viral particles comprising the rAAV vector, a pharmaceutical composition comprising the rAAV vector or the viral particles, and uses thereof in treating Glutaric aciduria type I (GA-I).

The application provides proline dehydrogenase, genes and mutants and applications thereof. The enzyme is derived from Pseudomonas xantholysinigenes and is recombinantly expressed in Escherichia coli. The target gene is codon-optimized when constructing a plasmid. The optimized DNA sequence is shown in SEQ ID NO. 2. The proline dehydrogenase is subjected to mutation modification. The enzyme and the mutants thereof can be used for efficiently catalyzing L-proline degradation to complete biological resolution of DL-proline, and high-purity D-proline is obtained. The application has great economic value in the field of green production of D-proline.

The invention relates to the technical field of mRNA vaccine research and development, and discloses a Tp0136T cell epitope mRNA vaccine based on lipid nanoparticle delivery and application of the Tp0136T cell epitope mRNA vaccine in syphilis prevention, the vaccine contains an mRNA sequence and an LNP delivery system, the mRNA sequence contains a Tp0136T1T cell epitope coding region (amino acid L477-S486, optimized by codon), a Cap 1 structure, optimized UTR and poly (A) tails of 65-76 adenosine, and psi or m5C is used for replacing trona; the LNP is prepared from SM-102, DSPC (Distearoyl Pyrrolidone), cholesterol and DMG-PEG (Dimethyl Glycol-Polyethylene Glycol) according to a molar ratio of 50 The particle size of the vaccine is less than or equal to 150nm, PDIlt; 0.2, the Zeta potential is-5 to-15 mV, and the encapsulation efficiency is greater than or equal to 93.47%. The preparation method comprises the steps of mRNA design synthesis, mRNA-LNP preparation characterization and in-vitro expression verification, through intramuscular injection inoculation, strong Th1 type and CD8 + CTL immune response can be induced, treponema pallidum load and skin ulcer rate can be reduced, and the method can be used for syphilis prevention.

The present disclosure provides pharmaceutical compositions containing nucleic acid molecules encoding PEDF which are codon-optimized, as well as methods for increasing the level of PEDF in the treatment or prophylaxis of retinal degenerative diseases, such as age-related macular degeneration (AMD), or other neurodegenerative diseases.

Provided are an expression cassette for AAV gene therapy of a neovascular ophthalmic disease, a plasmid for producing rAAV including the expression cassette, an rAAV produced by the plasmid, and use of the rAAV. By the expression cassette and the plasmid according to aspects, rAAV virus particles having reduced immunogenicity can be produced with high productivity and high quality, and an rAAV thus produced can reach a therapeutic concentration of aflibercept in the retina even at a low viral dose, thereby minimizing capsid-induced immune responses.

The invention discloses an Escherichia coli strain capable of stably expressing near-infrared fluorescent protein as well as a construction method and application of the Escherichia coli strain, and belongs to the technical field of gene engineering. According to the bacterial strain, escherichia coli probiotics Nissle 1917 serve as a chassis bacterial strain, at least two miRFP713 gene expression cassettes are integrated on a chromosome through a CRISPR-Cas mediated homologous recombination technology, and each expression cassette comprises a constitutive promoter and a miRFP713 gene optimized by a codon; meanwhile, the biliverdin synthetase gene is integrated to realize cofactor self-supply. The chromosome integrative Escherichia coli strain constructed by the invention realizes stable expression without exogenous induction, the fluorescence signal retention rate of 20 generations of passage is greater than 98%, and the fluorescence intensity is improved by 94.8 times, and is suitable for multiple application scenes such as long-term living body tracking, tumor targeted tracking and the like.

The invention discloses a human expression optimized Nipah virus N gene and application thereof in preparation of Nipah pseudovirus, belongs to the technical field of biology, and constructs a segment of Nipah virus N gene sequence optimized by human cell expression codons and constructs a recombinant vector pcDNA3.1-NiV-N-opti. The recombinant vector pcDNA3.1-NiV-N-opti is used as a pseudovirus expression vector, the recombinant vector pcDNA3.1-NiV-N-opti and pseudovirus packaging plasmids are used for preparing the kit for the nipah pseudovirus, the nipah pseudovirus can be efficiently obtained, the titer of the obtained nipah pseudovirus is higher than that of the nipah pseudovirus packaged by an original sequence, and an unexpected technical effect is achieved. Besides, the Nipah pseudovirus has His, Myc and EGFP marks, the preparation effect of the pseudovirus can be visually observed, and the Nipah pseudovirus can be used for Nipah serological diagnosis, virus neutralizing antibody evaluation and the like and has a good application prospect.

The invention belongs to the field of gene engineering, and particularly relates to application of chikungunya virus structural protein in improvement of pseudovirus stability, a construction method of the chikungunya virus structural protein and vaccines, antibody evaluation and in-vivo gene therapy products. The method comprises the following steps: constructing a human codon optimized Asian strain CHIKV-E3 + E2 + 6K + E1 structural protein eukaryotic vector, and designing two types of binder expression plasmids of targeted T cells; the vector, a psPAX2 helper plasmid and a lentivirus target vector containing an SFFV promoter and a ZsGreen reporter gene (or a CAR therapeutic gene) are co-transformed into a 293T cell, and the lentivirus is prepared through culture, filtration and centrifugation. The lentivirus can efficiently infect Jurkat cells and activate human T cells, obviously reduces the infection efficiency on 293T cells so as to reduce the off-target effect, has excellent stability in human serum, and provides a safe and efficient gene delivery tool for in-vivo CAR-T therapy.

The invention belongs to the technical field of veterinary biological products, and relates to an mRNA freeze-dried vaccine of a porcine epidemic diarrhea virus as well as a preparation method and application of the mRNA freeze-dried vaccine. When the mRNA vaccine is prepared, firstly, the S gene sequences of the porcine epidemic diarrhea virus GIIa type and the porcine epidemic diarrhea virus GIIc type which are separated and stored in a laboratory are selected; the preparation method comprises the following steps: firstly, carrying out codon optimization, then respectively connecting with 5 'UTR, 3' UTR, a termination codon and a polyA tail nucleic acid sequence in series, connecting to a vector, carrying out in-vitro template transcription, carrying out in-vitro transcription, capping and purifying to obtain mRNA, and finally, encapsulating and freeze-drying to prepare the mRNA vaccine. The invention further provides application of the vaccine to immunization of pigs, cellular immunity and humoral immunity in the pigs can be stimulated, the immunogenicity and protective efficacy of the vaccine are superior to those of commercially available bivalent live vaccines, and the vaccine can be used for preventing porcine diarrhea caused by infection of currently epidemic PEDV GIIa and GIIc type strains.

The invention discloses a heterologous expression pectate lyase engineering bacterium, a method for preparing pectate lyase by using the heterologous expression pectate lyase engineering bacterium and application of the heterologous expression pectate lyase engineering bacterium. Heterologous expression pectate lyase is recombinant protein optimized by codons, and the amino acid sequence of the heterologous expression pectate lyase is shown as SEQ ID NO.2; and the nucleotide sequence of the coding gene is as shown in SEQ ID NO.3. According to the invention, a recombinant expression vector containing the coding gene is constructed, pichia pastoris X-33 is used as host bacteria to construct engineering bacteria, and efficient expression of the pectate lyase is realized by optimizing vector elements and fermentation conditions. And the production time consumption is short, the cost is low, the process is easy to control, and the method is suitable for industrialization. And the produced enzyme has good purity, activity and stability, can efficiently degrade pectin substances when being used for tobacco, and improves the sensory quality and safety of cigarettes.

An isolated polynucleotide comprising a nucleotide sequence encoding a codon-optimised Complement Factor I (CFI).

The invention is characterized by a method and application of codon optimization of a gene product. Specifically, the present disclosure features a codon optimization method that promotes the increase in mRNA half-life and stability by reducing the frequency of m6A modification, thereby achieving the purpose of improving protein production. Additional methods of delivering codon optimized gene products are disclosed. The methods of the present disclosure are clinically related to gene and cell therapy as well as vaccine development.

The invention provides a sheep hepatic fascioliasis early specific diagnosis antigen FhCatL6 and a preparation method and application thereof, and relates to the field of immunological detection.The diagnosis antigen FhCatL6 is expressed based on BAA23743-1 gene, the diagnosis antigen FhCatL6 is recombinant protein FhCatL6, the amino acid sequence of the diagnosis antigen FhCatL6 is shown as SEQ ID NO.1, and the amino acid sequence of BAA23743-1 is shown as SEQ ID NO.2; according to the present invention, the sheep hepatic fascioliasis early-stage specific diagnosis antigen FhCatL6 is screened and determined, the antigen is expressed based on the BAA23743-1 gene, and is prepared by carrying out signal peptide cutting and codon optimization on BAA23743-1, such that the hepatic fascioliasis can be accurately identified, confusion with other similar diseases can be avoided, and the accuracy and the reliability of the diagnosis can be improved.

The invention discloses a recombinant bacillus calmette-guerin vaccine for expressing O and A type multi-epitope fusion peptides of foot and mouth disease viruses and application of the recombinant bacillus calmette-guerin vaccine, and belongs to the field of biotechnology and veterinary vaccines. The amino acid sequence of the multi-epitope fusion peptide is shown as SEQ ID NO: 1, the multi-epitope fusion peptide comprises dominant immune T cell and B cell antigen epitopes from a plurality of O-type and A-type foot-and-mouth disease virus epidemic strains, and a mycobacterium signal peptide is fused at the N terminal. The recombinant strain rBCG-MIPGA is successfully constructed by carrying out codon optimization on the fusion gene, cloning the fusion gene to a pMV306 vector and carrying out electrotransfection on BCG. The recombinant BCG can simultaneously excite high-titer specific antibodies aiming at foot-and-mouth disease virus type O and type A and remarkable T cell immune response, shows lasting and broad-spectrum protection potential, and has great application value in the aspect of preparing safe, efficient and broad-spectrum foot-and-mouth disease vaccines.

A self-inactivating lentiviral vector and method for treating or ameliorating Diamond-Blackfan anemia syndrome. The self-inactivating lentiviral vector includes nucleic acids encoding ribosomal protein S19 operably linked to an EF1α promoter or a MND promoter, wherein the nucleic acids encoding RPS19 are codon optimized for expression in human cells and are devoid of aberrant splice acceptor sites, thereby improving the safety and efficacy of the lentiviral vector.

What is disclosed herein are nucleic acids encoding a chimeric fusion protein comprising an extracellular domain that comprises an antigen binding domain, particularly a TROP2 binding domain. The nucleic acids disclosed here comprise codon-optimized coding sequence for the chimeric fusion protein and comprises a 5' UTR and a 3' UTR, wherein the 3' UTR comprises at least one microRNA binding site.

The invention relates to the technical field of veterinary drugs, and particularly provides a polynucleotide molecule composition for preventing cat infectious peritonitis, a chimeric polynucleotide molecule and an mRNA vaccine. In order to overcome the ADE effect caused by S protein full-length antigen in the prior art, FIPV N protein without ADE risk is adopted and is combined with screened S protein specific T cell and B cell epitopes. Core schemes include two classes: one is a separate polynucleotide composition comprising epitope peptides encoding N and S proteins; and 2, a single chimeric polynucleotide encoding N protein and S protein epitope peptides. The polynucleotides are optimized by codons and encapsulated in lipid nanoparticles (LNP) to prepare vaccines. Experiments prove that the vaccine can synergistically stimulate powerful immune response, the survival rate of immune cats is remarkably increased to 80%-100% from standard 40% on the premise that ADE is not caused, and FIP is effectively prevented.

The invention relates to a method for heterologously producing ganoderic acid, which comprises the following steps: respectively transforming candidate oxidase, candidate reductase and candidate acetyltransferase into corresponding saccharomyces cerevisiae strains for heterologously expressing, and screening the transformed strains through intracellular fermentation or whole-cell catalysis to obtain codon-optimized AfuSDR and CsSDR capable of catalyzing oxidization of hydroxyl of a ganoderma triterpene skeleton C3, the method comprises the following steps of: performing independent expression or combined expression on reductases AKR1C2, AKR1C4 and FusC1 which are optimized by codons and are used for catalyzing C3 keto reduction and acetyltransferase BsAT which is optimized by codons and is used for catalyzing C3 hydroxyl acetylation, so as to realize heterologous biosynthesis of ganoderic acid TR (GA-TR), gibberellic acid Ja (GA-Ja) and ganoderic acid Mf (GA-Mf).