187 results about "Codon optimized" patented technology

The present invention belongs to the field of biology, and relates to a nucleic acid molecule encoding an anti-NKG2A antibody, a recombinant vector comprising the nucleic acid molecule, and an expression method wherein the nucleic acid molecule comprises codons optimized for expression cells. By utilizing the nucleic acid molecule disclosed by the invention, the expression quantity of the anti-NKG2A antibody in cells can be increased, the yield of the anti-NKG2A antibody is increased, and the production cost is reduced.

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 invention provides a method and a device for designing a protein coding codon sequence based on a large multi-modal model, which comprehensively utilize information such as an amino acid sequence, space folding and mRNA quality of protein to realize high-quality, controllable and evaluable codon sequence design. According to the method, the dependence of a traditional codon optimization strategy on a reference gene template is broken through, and a codon optimization problem is converted into a codon design task based on protein information. Through comprehensive consideration of information such as space folding of protein, context of amino acid sequences, mRNA quality and the like, the model can generate a codon sequence which is more in line with codon use preference of specific species, reasonable in structure and efficient in expression, so that a new thought and technical support are provided for safe, stable, efficient and controllable protein expression.

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 relates to bacillus subtilis for expressing bovine-derived beta-lactoglobulin and a construction method of the bacillus subtilis, and belongs to the technical field of microorganisms. According to the bacillus subtilis provided by the invention, bacillus subtilis BSF01 is used as a host, a constitutive promoter P566 is used for heterologous expression of a bovine-derived beta-lactoglobulin coding gene optimized by a codon, the yield of beta-lactoglobulin obtained by shake flask fermentation for 30 hours reaches 0.37 g / L, the yield of beta-lactoglobulin obtained by 3L fermentation tank fermentation for 44 hours reaches 3.06 g / L, the production intensity is 0.07 g / L / h, and the production cost is low. The bovine-derived beta-lactoglobulin is efficiently and insolubly expressed in a food safety level strain, the food industrial application of the beta-lactoglobulin is promoted, and the problems of food safety and excessive glycosylation possibly existing in heterologous expression of the beta-lactoglobulin in the prior art are solved.

The invention discloses a streptomyces mini-gene editing system based on coupled TnpB and reconstructed NHEJ as well as a construction method and application of the streptomyces mini-gene editing system. The system comprises TnpB nuclease which can be expressed in prokaryotic microorganisms through codon optimization, a reRNA guide box of the TnpB nuclease, and a LigD gene expression box, the prokaryotic microorganism itself does not have NHEJ or has a defective NHEJ pathway (such as streptomyces); the reRNA guide box is used for guiding TnpB protein to move towards a target sequence; and the LigD gene expression cassette is used for expressing ligase LigD and reconstructing the prokaryotic microorganism DNA repair system NHEJ. According to the system disclosed by the invention, small InDels can be efficiently introduced near a target site under the action of a reconstructed streptomyces repair system, so that editing of a target gene is realized, and a gene large fragment deletion phenomenon introduced by self-defect repair is avoided.

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.

Provided herein are codon-optimized Smad7 polynucleotides and vectors comprising codon-optimized Smad7 polynucleotides for use in increasing or prolonging Smad7 expression in a subject.

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 belongs to the technical field of microorganisms, and particularly relates to a method for expressing human lactoferrin by using lactococcus lactis. The method comprises the following steps: step 1, obtaining a codon-optimized hLF gene nucleotide sequence; 2, constructing a recombinant plasmid containing the hLF gene nucleotide sequence obtained in the step 1; and step 3, transferring the recombinant plasmid obtained in the step 2 into lactococcus lactis for induced expression. The heterologous expression of the human lactoferrin hLF is realized by utilizing the lactococcus lactis, the method is low in cost, convenient and fast, and a new way is provided for high-purity and high-yield preparation of the human lactoferrin hLF.

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 invention provides antigen protein HPA of bovine-derived A-type pasteurella multocida and a preparation method and application of recombinant protein of the antigen protein HPA. The amino acid sequence of the antigen protein HPA is shown as SEQ ID NO.3. The preparation method comprises the following steps: cloning a coding gene sequence, shown as SEQ ID NO.1, of the antigen protein HPA after codon optimization into a prokaryotic expression vector; a recombinant plasmid containing the antigen protein HPA coding gene is obtained; transforming the recombinant plasmid into escherichia coli competent cells to obtain a recombinant expression strain; the antigen protein HPA is obtained through fermentation culture, inducible expression and purification. According to the invention, mice are immunized, the protection rate can reach 100%, and the problem of poor protection effect of the A-type pasteurella subunit vaccine for bovine pneumonia at present is solved.

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 provides circular polyribonucleotides including a sequence including: a circularization element; a first spacer; an internal ribosome entry site (IRES) sequence; a sequence encoding a polypeptide; and a second spacer, where the sequence encoding the polypeptide has been codon-optimized to reduce the number of uracil ribonucleotides and / or reduce or remove TLR7- and TLR8-recognition motifs, and kits and compositions thereof. The present disclosure also provides methods of treating subjects in need thereof with the circular polyribonucleotides and compositions described herein.

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 present disclosure provides a construction method of a bacterial cellulose (BC)-enriched plant using multi-gene tandem and use thereof, and relates to the technical field of genetic engineering. In the present disclosure, an acsAB gene, an acsC gene, and an acsD gene are combined and then subjected to codon optimization according to a codon preference of the crop. Resulting optimized genes are fused with a 35S promoter and a nopaline synthase (NOS) terminator to obtain three gene expression cassettes. The three gene expression cassettes are ligated into a plant expression vector to obtain a multi-gene plant transformation vector containing the above three gene expression cassettes. The multi-gene plant transformation vector is then transformed into rice to obtain a transgenic rice plant capable of synthesizing the BC. It is determined that the transgenic rice plant has a BC content of 3.81%.

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.