334 results about "Codon optimized" patented technology

The invention provides a high-stability recombinant humanized elastin, a genetically engineered bacterium for expressing the protein and application of the high-stability recombinant humanized elastin, and belongs to the technical field of binding tissue peptides. The nucleotide sequence of the recombinant humanized elastin is shown as SEQ ID NO.1; and the amino acid sequence is as shown in SEQ ID NO.4. According to the invention, the nucleotide sequence of the humanized elastin is subjected to codon optimization and transferred into escherichia coli to construct the genetically engineered bacterium for expressing the recombinant humanized elastin, so that the expression quantity of the recombinant humanized elastin can be increased, the stability of the recombinant humanized elastin can be improved, and the recombinant humanized elastin can be used for preparing the recombinant humanized elastin. The migration rate of the recombinant humanized elastin to the HSF cell is improved, and the expression quantity of the h-ELN gene, the h-COL1A1 gene and the h-COL3A1 gene of the HSF cell is improved.

Provided herein are methods and compositions for expressing Otoferlin, e.g., utilizing adeno-associated viral (AAV) particles. Further provided herein are compositions of AAV particles comprising one or more polynucleotides encoding Otoferlin are codon optimized for expression in human cells. Such methods and compositions may be useful for treatment of diseases and disorders such as Deafness, Autosomal Recessive 9 (DFNB9). Also provided are kits comprising such compositions.

The invention discloses an OpenCRISPR (Open Clustered Regularly Interspaced Short Palindromic Repeats)-NLS (Non-Least Squares) fusion protein for gene editing of penaeus monodon, a construction method of the OpenCRISPR-NLS fusion protein and a nucleotide construct containing the OpenCRISPR The invention also discloses an application of the fusion protein or the nucleotide construct in gene editing of penaeus monodon and an application of the fusion protein or the nucleotide construct in population improvement of aquaculture penaeus monodon. According to the invention, codon optimization is carried out on a coding sequence of OpenCRISPR-1 protein, and a nuclear localization signal (NLS) is fused with the OpenCRISPR protein subjected to codon optimization; the method has the advantages of being efficient in gene expression, accurate in targeting positioning capacity, excellent in genome editing efficiency, capable of reducing the off-target effect, wide in adaptability, capable of promoting sustainable aquaculture, remarkable in economic and social value and the like.

The invention relates to the technical field of microbial genetic engineering, in particular to a method for improving cordycepin synthesized by saccharomyces cerevisiae and saccharomyces cerevisiae engineering bacteria. According to the invention, codon-optimized 2 '-carbonyl-3'-deoxyadenosine reductase gene and 3 '-adenosine monophosphate phosphohydrolase gene from cordyceps militaris are expressed by using plasmids, and then the plasmids are transformed into host cells to obtain the saccharomyces cerevisiae engineering bacteria. And inoculating a seed solution obtained by seed culture of the activated saccharomyces cerevisiae engineering bacteria into a fermentation culture medium containing a metabolic effector, and carrying out fermentation culture to synthesize cordycepin. Metabolic effectors (Cu < 2 + >, Fe < 2 + >, Mg < 2 + >, Zn < 2 + >, citric acid, cysteine, aspartic acid, glycine, VB1 and tea polyphenol) are added into a fermentation culture medium strain, and the variety and concentration of the metabolic effectors are optimized, so that the yield and synthesis efficiency of cordycepin are remarkably improved. By using the combination of the strain obtained by the invention and the optimal effector, the yield of cordycepin in a fermentation tank reaches 2.9 g / L.

Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. The disclosure provides fusion proteins of nucleic acid programmable DNA binding proteins (napDNAbp), e.g., Cas9 or variants thereof, and nucleic acid editing proteins such as cytidine deaminase domains (e.g., novel cytidine deaminases generated by ancestral sequence reconstruction), and adenosine deaminases that deaminate adenine in DNA. Aspects of the disclosure relate to fusion proteins (e.g., base editors) that have improved expression and / or localize efficiently to the nucleus. In some embodiments, base editors are codon optimized for expression in mammalian cells. In some embodiments, base editors include multiple nuclear localization sequences (e.g., bipartite NLSs), e.g., at least two NLSs. In some embodiments, methods for targeted nucleic acid editing are provided.

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 present invention provides codon optimized Factor IX sequences, vectors and host cells comprising codon optimized Factor IX sequences, polypeptides encoded by codon optimized Factor IX sequences, and methods of producing such polypeptides. The present invention also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject a codon optimized Factor IX nucleic acid sequence or the polypeptide encoded thereby.

Disclosed are AAV viral-based vector compositions useful in delivering a variety of nucleic acid segments, including those encoding therapeutic polypeptides to selected mammalian host cells for use in therapeutic autoimmune modalities, including, for example, the in vivo induction of immunological tolerance via a liver-directed AAV-based gene therapeutic regimen for treating and / or ameliorating autoimmune disorders such as multiple sclerosis. Further disclosed are nucleic acid segments encoding therapeutic polypeptides that have been codon-optimized for expression in human cells.

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 discloses a genetically engineered algae for inhibiting protozoa and a construction method and application thereof, and is characterized in that the genetically engineered algae is phaeodactylum tricornutum into which an expression plasmid pPha-T1-eGFP-Linker-EP2 is introduced, and the nucleotide sequence of the expression plasmid is as shown in SEQ ID NO: 7. The construction method comprises the following steps: artificially synthesizing an EP2 coding sequence optimized by a codon; a step of inserting the expression vector into the downstream of the eGFP of the expression vector, and expressing a plasmid pPha-T1-eGFP-Linker-EP2; the method comprises the following steps of: electrically transforming expression plasmids into phaeodactylum tricornutum, and coating transformed phaeodactylum tricornutum cells on an f / 2 plate culture medium for culturing and screening positive algae strains to obtain the genetically engineered algae for inhibiting protozoa, and has the advantages that the large-scale culture efficiency and success rate of microalgae are improved, and the genetically engineered algae have a strong inhibition effect on Tophelenium sectorius and Tophelenium lakeri.

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.

According to the recombinant human VII type collagen as well as the preparation method and the application thereof, nucleotide optimized by codons is recombined into plasmids, and the plasmids are expressed in host cells, so that the recombinant human VII type collagen can be efficiently obtained; the recombinant human VII type collagen has collagen activity for promoting cell proliferation and cell adhesion, can inhibit metalloproteinase expression, has a cell repair effect, and can be used as a cell repair product; the method can be widely applied to food, cosmetics, health care products or medical equipment products.

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 discloses a flocculation-controllable genetic engineering alga and a construction method and application thereof, and is characterized in that the genetic engineering alga is phaeodactylum tricornutum into which an enabling plasmid pPha-J47869-EG01992-eGFP-FSP is introduced, the FSP is directionally expressed on the outer side of a phaeodactylum tricornutum cell, and the construction method comprises the following steps: constructing an induced expression plasmid pPha-J47869-eGFP and a directional expression plasmid pPha-J47869-EG01992-eGFP; artificially synthesizing a codon-optimized FSP coding sequence, inserting the codon-optimized FSP coding sequence into the downstream of the eGFP of the directional expression vector, and constructing an enabling plasmid; finally, the enabling plasmids are electrically transformed into phaeodactylum tricornutum, an f / 2 plate culture medium is coated with the phaeodactylum tricornutum for culture, positive algae strains are screened and identified, and the gene engineering algae with controllable flocculation are constructed and have the advantages that the gene engineering algae can be efficiently combined with white carbon black, and the enabling process of microalgae is efficiently coupled with the culture and collection process of the microalgae.

The invention discloses a method for creating melatonin-rich soybeans by utilizing synthetic regulatory elements and application of the melatonin-rich soybeans. According to the invention, a soybean standard line Williams 82 (W82) is used as a receptor, melatonin pathway genes COMT and SNAT are optimized through codons to synthesize regulation elements to efficiently and accurately activate expression of COMT and SNAT, and specific accumulation in soybean seeds is realized, so that a new soybean material rich in melatonin in the seeds is created. Compared with a control receptor, the melatonin content of the modified soybean is increased by more than 30 times. The method for efficiently producing the melatonin by synthesizing the regulatory element has an important value for increasing the content of the melatonin, provides a new thought and tool for improving the quality of soybeans and crops, is beneficial for supplementing high-value active substances in daily diet, and has a wide application prospect. The method has strong application potential in the aspect of assisting biosynthesis of plant-derived natural products.

The invention discloses a peste des petits ruminants virus specific monoclonal antibody and application thereof.The monoclonal antibody is obtained by screening peste des petits ruminants virus N protein, the amino acid sequence of the peste des petits ruminants virus N protein is shown as SEQ ID NO: 1, the peste des petits ruminants virus N protein is obtained by transcription of a peste des petits ruminants virus N gene, and the peste des petits ruminants virus N gene is shown as SEQ ID NO: 2. The nucleotide sequence of the codon-optimized peste des petits ruminants virus N gene is shown as SEQ ID NO: 2. The monoclonal antibody provided by the invention can be specifically combined with the peste des petits ruminants virus N protein, the titer is high, the affinity is good, meanwhile, the detection kit prepared based on the antibody pair can effectively carry out peste des petits ruminants virus detection, and a technical support can be provided for epidemiological monitoring and diagnosis of peste des petits ruminants virus infection.

The invention discloses recombinant human chemical fibronectin as well as a coding gene and application thereof. The method comprises the following steps: performing codon optimization on an amino acid sequence of recombinant human chemical fibronectin according to codon preference of pichia pastoris to obtain a coding gene nucleotide sequence SEQ ID NO.2, cloning the coding gene nucleotide sequence SEQ ID NO.2 between EcoRI and NotI restriction enzyme cutting sites of a pichia pastoris expression vector pPIC9K to obtain a recombinant expression vector pPIC9K-FNc, and transferring the recombinant expression vector pPIC9K-FNc into a P.pastoris GS115 expression host cell to obtain the recombinant human chemical fibronectin. Recombinant pichia pastoris P.pastoris GS115 / pPIC9K-FNc is obtained through genetic mycin G418 screening, then recombinant bacteria are subjected to fermentation culture and are subjected to induced expression by using methanol, and then the recombinant human chemical fibronectin is obtained after purification. The fibronectin produced by the method is large in molecular weight and has better biological activities such as wrinkle resistance and barrier repair.

To provide recombinant adeno-associated viral vectors for treating Bietti Crystalline Dystrophy.SOLUTION: Provided are a recombinant adeno-associated vector comprising a codon-optimized sequence encoding CYP4 V2 linked to predetermine gene expression regulatory sequences, and its use in treating Bietti Crystalline Dystrophy (BCD).SELECTED DRAWING: None

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 flocculation-controllable genetic engineering alga and a construction method and application thereof, and is characterized in that the genetic engineering alga is phaeodactylum tricornutum into which an enabling plasmid pPha-J47869-EG01992-eGFP-FSP is introduced, the FSP is directionally expressed on the outer side of a phaeodactylum tricornutum cell, and the construction method comprises the following steps: constructing an induced expression plasmid pPha-J47869-eGFP and a directional expression plasmid pPha-J47869-EG01992-eGFP; artificially synthesizing a codon-optimized FSP coding sequence, inserting the codon-optimized FSP coding sequence into the downstream of the eGFP of the directional expression vector, and constructing an enabling plasmid; finally, the enabling plasmids are electrically transformed into phaeodactylum tricornutum, an f / 2 plate culture medium is coated with the phaeodactylum tricornutum for culture, positive algae strains are screened and identified, and the gene engineering algae with controllable flocculation are constructed and have the advantages that the gene engineering algae can be efficiently combined with white carbon black, and the enabling process of microalgae is efficiently coupled with the culture and collection process of the microalgae.

The present invention provides codon optimized Factor IX sequences, vectors and host cells comprising codon optimized Factor IX sequences, polypeptides encoded by codon optimized Factor IX sequences, and methods of producing such polypeptides. The present invention also provides methods of treating bleeding disorders such as hemophilia comprising administering to the subject a codon optimized Factor IX nucleic acid sequence or the polypeptide encoded thereby.

The invention discloses an mRNA (messenger ribonucleic acid) vaccine for preventing and / or treating pseudorabies and application of the mRNA vaccine. Codon optimization is performed on a gene sequence of a pseudorabies virus gD protein antigen to obtain an optimized gene sequence of the pseudorabies virus gD protein antigen; the pseudorabies virus truncated gD protein antigen is obtained by optimizing the sequence of the pseudorabies virus gD protein antigen, the gene sequence of the truncated gD protein antigen is subjected to codon optimization, and experiments show that the optimized pseudorabies virus truncated gD protein antigen has higher expression quantity; the sequence-optimized pseudorabies virus truncated gD protein is further prepared into an mRNA vaccine, and after the mRNA vaccine is used for immunizing mice and piglets, it is found that the mRNA vaccine of the sgD protein can play an immune protection role on the mice and the piglets, and the mRNA vaccine is obviously superior to commercially available pseudorabies inactivated vaccines.

The present disclosure provides exemplary codon optimized nucleic acid sequences encoding Piscine Myocarditis Virus (PMCV). Expression vectors comprising one or more of the described codon optimized nucleic acids sequences are also provided.

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.

A codon-optimized polynucleotides of ATP7B and truncated ATP7B. It further relates to expression constructs, vectors, viral particles, or compositions comprising the polynucleotide of the disclosure, and methods and uses of the polynucleotides, expression construct, vectors, viral particles, or compositions in gene therapy for treating Wilson disease (WD).

Disclosed herein is a method of treating Pincare Disease, the method comprising administering a recombinant AAV (rAAV) vector comprising an rAVV genome comprising a heterologous nucleic acid encoding a GAA signal peptide or portion thereof, a heterologous signal peptide and an acidic alpha-glucosidase (GAA) polypeptide, or an N-terminal truncation thereof, the heterologous nucleic acid is operably linked to a liver-specific promoter wherein the nucleic acid encoding the GAA polypeptide may be a wild-type nucleic acid sequence, or a modified nucleic acid sequence, or a codon-optimized nucleic acid sequence, or a modified nucleic acid sequence, or a codon-optimized nucleic acid sequence, or a modified nucleic acid sequence, or a codon-optimized nucleic acid sequence. And may optionally be modified to reduce or completely eliminate CG and CpG dinucleotides and optionally eliminate alternative reading frame (ARF) content.

The invention relates to application of a gene pmtA in improving broad-spectrum tolerance of escherichia coli. According to the present invention, the research results show that the codon-optimized (derived from Rhodobacter sphaeroids) gene pmtA for coding phosphatidyl ethanolamine-N-methyltransferase EC 2.1. 1.17 is integrated to the escherichia coli genome ldhA site, such that phosphatidylcholine can be introduced to remodel the escherichia coli cell membrane phospholipid system so as to improve the broad-spectrum tolerance of escherichia coli; in addition, an artificial regulation element M1-12 and a gene pmtA are assembled to construct a tolerance module, and the tolerance module is integrated to a lactic dehydrogenase encoding gene ldhA site of an escherichia coli genome through a lambda Red homologous recombination method to obtain an engineering bacterium named + PC. According to the engineering strain provided by the invention, the broad-spectrum tolerance of the strain is improved, the methanol utilization capability of escherichia coli is remarkably enhanced while the tolerance of an intermediate toxic substrate formaldehyde is enhanced, and the TAL synthesis capability is remarkably enhanced while the tolerance to TAL is enhanced.

The present invention provides: a codon-optimized polynucleotide encoding factor IX (FIX); a recombinant AAV-producing plasmid comprising the polynucleotide; a recombinant AAV produced by the plasmid; and use of the recombinant AAV for treating hemophilia B. The plasmid comprising the polynucleotide can produce a recombinant AVV that exhibits high FIX expression and activity and has excellent productivity and high quality.