280 results about "Codon optimization" patented technology

The invention discloses a recombinant humanized XVII type collagen as well as an expression vector and a genetically engineered bacterium thereof. Four functional fragments with high activity are obtained through screening, one of the fragments is optimized and then combined according to a certain sequence to obtain recombinant humanized XVII type collagen JYC1703, or one of the fragments is optimized and then combined according to a certain sequence, and GPP sequences are fused at an amino terminal and a carboxyl terminal respectively to obtain recombinant humanized XVII type collagen JYC1701 and JYC1702, and the recombinant humanized XVII type collagen JYC1701 and JYC1702 are combined according to a certain sequence to obtain the recombinant humanized XVII type collagen JYC1701 and JYC1702. After codon optimization, amplifying and cloning into an expression vector, transforming into pichia pastoris competent cells after enzyme digestion linearization, and screening high-copy recombinants through G418 resistance gradient, so as to obtain pichia pastoris genetically engineered bacteria for expressing recombinant humanized XVII type collagen. The recombinant humanized XVII type collagen JYC1701, JYC1702 and JYC1703 disclosed by the invention show excellent functions of promoting cell adhesion, cell proliferation and cell migration, and are suitable for preparing products such as skin care products or medical instruments and the like.

The invention relates to a saccharomyces cerevisiae strain for efficiently synthesizing cordycepin as well as a construction method and application of the saccharomyces cerevisiae strain, and belongs to the technical field of microbial genetic engineering. The construction method provided by the invention comprises the following steps: carrying out multi-copy genome integrated expression on heterologous codon optimized 2 '-carbonyl-3'-deoxyadenosine reductase genes and 3 '-adenosine monophosphate phosphohydrolase genes, so as to increase the dosage of key enzymes; and supplementing the defective gene of the engineering strain to obtain the saccharomyces cerevisiae strain B5U3 with high yield of cordycepin. According to the obtained saccharomyces cerevisiae strain for efficiently synthesizing the cordycepin, the cordycepin yield reaches 2.92 g / L after shake flask fermentation, the cordycepin yield reaches 7.5 g / L after fermentation in a fermentation tank, and a foundation is laid for safer and more reliable industrial production of the cordycepin.

The invention discloses application of a gene FTO in improvement of forage grass quality, yield and stress resistance. According to the invention, the FTO gene is subjected to codon optimization according to the metatropism of alfalfa and is overexpressed in alfalfa to obtain a transgenic alfalfa plant overexpressed with the FTO gene, and compared with wild alfalfa, the transgenic alfalfa plant overexpressed with the FTO gene has the advantages that the growth performance is enhanced; the crude protein content, the soluble carbohydrate content and the in-vitro dry matter digestibility are obviously higher than those of the wild type; the crude fiber content, the acid detergent fiber content, the neutral detergent fiber content and the lignin content of the transgenic alfalfa plant are obviously lower than those of a wild type; in addition, the drought resistance and salt stress resistance of the transgenic alfalfa plant are also obviously enhanced, which indicates that the FTO gene has the functions of improving forage grass quality, resisting stress or positively regulating plant growth. The method has application prospects in the aspects of improving forage grass quality and stress resistance or promoting plant growth and the like.

A technique related to codon optimization is provided. In a method for optimizing nucleic acid sequences expressing proteins in a host, protein subsequence-nucleic acid subsequence pairs are obtained from collected highly expressed protein sequences and their encoding nucleic acid sequences to form a training set. And training a neural machine translation model by using the training set. The neural machine translation model realizes translation from an amino acid sequence to a codon sequence. A protein sequence needing codon optimization is cut into protein subsequences. And translating a protein subsequence from an amino acid sequence to a codon sequence by using the trained neural machine translation model. The translated subsequences are superimposed to merge into a full-length codon sequence. In the process of superimposing to synthesize the codon sequence, according to the frequency or number of synonymous codons corresponding to each amino acid position of the protein sequence, the synonymous codon with the highest occurrence frequency or number is selected as the optimal codon at the position.

The invention discloses a grass carp gamma interferon fusion protein as well as a mutant, a coding gene and application thereof. In order to improve the expression quantity or expression efficiency of the grass carp gamma interferon, codon optimization and mutation are carried out on the coding gene of the grass carp gamma interferon to obtain the grass carp gamma interferon mutant. The fusion protein is further obtained by connecting the grass carp gamma interferon mutant with the C end of the grass carp ferritin heavy chain subunit of which the last 17 amino acids are removed through a connecting peptide. In order to further improve the antiviral activity of the fusion protein, the obtained fusion protein is subjected to single-point mutation, and the titer of the fusion protein is remarkably improved. The fusion protein is expressed by a silkworm cell eukaryotic expression system, and the fusion protein shows a conformation suitable for the interferon to play functions based on the ferritin self-assembly characteristic, so that the titer of the interferon in a host is effectively improved, and the in-vivo and in-vitro half-life period of the interferon is prolonged. The invention has application prospects in preparation of drugs or reagents for preventing or treating viral diseases of grass carp and the like.

The invention relates to the field of prevention and treatment of respiratory diseases, and discloses a preparation method of a human respiratory syncytial virus preventive mRNA vaccine. The main components of the mRNA vaccine comprise mRNA for coding an antigen and lipid nanoparticles. According to the mRNA sequence provided by the invention, an independently researched and developed 5 'UTR / 3' UTR sequence and a codon optimization mode are adopted, and efficient translation of target protein can be realized. According to the mRNA vaccine provided by the invention, RSV F protein is selected as an antigen, and the RSV F protein sequence is optimally designed, so that stable pre-fusion conformation RSV F protein can be expressed, and after a mouse is immunized, the mouse can be induced to generate high-level RSV F protein specific binding antibody titer and neutralizing antibody titer.

The invention relates to the technical field of gene engineering, in particular to nucleic acid for encoding kreotoxin and a preparation method of the kreotoxin. The invention provides nucleic acid for encoding the botulinum toxin, the nucleic acid is subjected to codon optimization, and the scheme is proved to be superior to other optimized sequences. Through verification, the sequence has a relatively high expression quantity in escherichia coli and is in soluble expression, and the prepared kreotoxin has good activity.

The invention belongs to the technical field of bioengineering, and provides a recombinant XVII type collagen as well as a preparation method and application thereof. A target gene sequence is obtained through bioinformatics predictive analysis and integrated design on the basis of a human XVII type collagen sequence, codon optimization is carried out, a T7 promoter in a plasmid vector is optimized, an engineering strain of the high-hydrophobicity humanized XVII type collagen is prepared, and the high-hydrophobicity humanized XVII type collagen is obtained. The strain is subjected to culture and inducible expression to obtain high-yield recombinant XVII type collagen. The recombinant XVII type collagen disclosed by the invention is reasonable in sequence design, large in expression quantity, simple in preparation method and stable in heredity, and rapid separation of the collagen can be realized in a production and preparation process by utilizing high hydrophobicity.

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

The invention discloses a method for fermenting recombinant humanized collagen by using escherichia coli and application of the recombinant humanized collagen, and belongs to the field of collagen. The amino acid sequence of the prepared recombinant humanized III-type collagen is shown as SEQ ID NO.4-7, the recombinant humanized III-type collagen is formed by connecting a plurality of repetitive units of natural collagen in series and has the advantages of being good in biocompatibility and high in safety, in addition, the nucleotide sequence for coding the recombinant collagen is designed according to the codon optimization theory, and the recombinant humanized III-type collagen has the advantages of being good in biocompatibility and high in safety. And carrying out fermentation expression on escherichia coli to obtain the recombinant humanized III-type collagen. The humanized III-type collagen can significantly improve the cell adhesion activity and proliferation activity, and can be widely applied to the fields of food, cosmetics, health care products, medical instruments and the like.

The invention relates to a codon optimization method for improving the protein expression quantity of a target gene in a plant, and belongs to the technical field of gene engineering. The invention provides a codon optimization method for improving the protein expression quantity of a target gene in a plant, which comprises the step of replacing the third basic group of a codon of a target protein coding sequence with C or G to obtain an optimized target protein coding sequence. The method has effectiveness and universality in plants, and research shows that after codon optimization of the method, the protein expression quantity of target genes in a plant protein transient expression system and a transgenic plant is increased by several times to dozens of times. According to the production application of the functional protein taking the plant as the bioreactor, the yield of the target protein can be increased, and the protein purification cost can be reduced; according to the application of the method in preparation of the transgenic plant, the expression quantity of the target gene can be increased, so that the target character of the transgenic plant is enhanced, and the screening cost of the high-expression transgenic plant is reduced.

The invention provides a fusion protein for promoting proliferation of hair follicle stem cells as well as a preparation method and application of the fusion protein. The fusion protein is formed by fusion expression of a plurality of collagen structural domains capable of being combined with cell integrin and / or specific structural domains from certain functional proteins screened from collagen XVII and III type collagen. Nucleic acid molecules for coding the fusion protein are subjected to codon optimization and then recombined and stably expressed in escherichia coli to obtain the fusion protein. The fusion protein has excellent performance of promoting proliferation and migration of human hair follicle stem cells and human fibroblasts, can be used for repairing skin and hair follicle tissues and improving resistance of skin and hair follicles, and is suitable for popularization and application in the fields of beauty and skin care, medical equipment, tissue engineering and the like.

The invention belongs to the technical field of biology, and discloses a preparation method of fusion protein of salmonella typhimurium flagellin and Seneca virus antigen. Escherichia coli Rosetta (DE3) is selected as a host, a salmonella typhimurium flagellin gene and a Seneca virus antigen gene are fused and are cloned to a pET-28a (+) vector after codon optimization, the obtained fusion protein keeps the flagellin adjuvant activity, the solubility and the expression level of VP2 are improved, the yield can reach 40 mg / L, and the fusion protein can be used for preparing a recombinant vector. The expression and purification process cost is greatly reduced. The fusion protein provided by the invention can activate a TLR5 pathway and mouse macrophages, is simple in production process and low in cost, and has important significance on research and production of Seneca virus subunit vaccines.

The invention discloses an echinococcosis mRNA (messenger Ribonucleic Acid) vaccine as well as a preparation method and application thereof. The preparation method of the mRNA vaccine for the echinococcosis comprises the following steps: carrying out codon optimization on a modified target antigen protein, then assembling the modified target antigen protein with 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a pUC57 plasmid, and sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmid to prepare an mRNA molecule, thereby obtaining the mRNA vaccine for the echinococcosis. Finally, mRNA molecules are wrapped in lipid nanoparticles through a microfluidic method to form the mRNA vaccine for the echinococcosis, and immune effect evaluation is carried out on the mRNA vaccine for the echinococcosis. Experiments prove that the prepared mRNA vaccine for the echinococcosis can activate humoral immunity and cellular immunity of mice at the same time, can provide an effective protection effect for the mice attacking insects, and has a wide application prospect in the aspect of preventing and / or treating the echinococcosis.

The invention relates to the technical field of collagen, and discloses a preparation method of recombinant engineering bacteria for expressing III-type collagen COL17A1, which comprises the following steps: synthesizing an optimized sequence of human III-type collagen, and adding proper restriction enzyme cutting sites at two ends of the sequence. According to the preparation method of the recombinant engineering bacterium for expressing the III-type collagen COL17A1, codon optimization is carried out on a target sequence by adopting a yeast expression system, OptimGeneTM codon optimization is provided according to codon preference of yeast strains such as pichia pastoris, and the yield of recombinant protein is further improved. And a proper vector is selected to realize high-efficiency expression in pichia pastoris. For example, a pGAPZ A-alpha A vector stably expresses recombinant protein in pichia pastoris by using a GAP promoter, is a constitutive gene expression vector, and can maintain a stable expression rate in cells no matter how the environment changes. An inducer does not need to be added, the production process is simplified, the production cost is reduced, and the production efficiency is improved.

The invention relates to the technical field of molecular biology, in particular to a codon optimization method. The invention provides a codon optimization method, the codon is replaced according to parameters such as codon type and quantity, local GC content, local repetitive sequence, mRNA secondary structure, mRNA free energy and the like, and the optimized sequence can be subjected to experimental screening to obtain a DNA sequence of high-expression protein. The downstream of the method is mainly the application in the IVT mRNA direction: the optimized DNA sequence is constructed into a vector, the plasmid is linearized to serve as a template for in-vitro mRNA transcription, the mRNA is encapsulated into LNP or LNP coupled with other molecules, and the protein expression in 293T cells (not limited to 293T cells) or mice is obviously improved.

The invention discloses a construction method and application of a chlamydomonas reinhardtii-sourced high-sweetness protein variant engineering algal strain, the construction method comprises the following steps: carrying out gene modification on sweeteners Thaumatin and Brazzein, optimizing preference codons of chlamydomonas reinhardtii to synthesize CrThaumatin and CrBrazzein encoding genes, cloning the encoding genes into an expression vector pGM6, introducing the recombinant expression vector into a chlamydomonas reinhardtii wild-type chlamydomonas sp. Algal strain, and carrying out high-sweetness protein variant engineering algal strain of the chlamydomonas reinhardtii-sourced high-sweetness protein variant engineering algal strain of the chlamydomonas reinhardtii. And screening by using a paromomycin resistance marker carried by the expression vector to successfully obtain an engineering algal strain. According to the present invention, the engineering strain is subjected to fermentation production to obtain the dry and stable Chlamydomonas reinhardtii powder rich in the target sweet protein, and the protein purification oral test results show that the sweet taste of the Chlamydomonas reinhardtii powder is 3-5 times of the sweet taste of the natural Thaumatin and Brazzein protein;

The invention discloses a codon optimization method and device, equipment and a storage medium, and the method comprises the steps: obtaining a target protein sequence; inputting the target protein sequence into a pre-trained target codon optimization model to obtain a codon optimization result of the target protein sequence; wherein the target codon optimization model comprises at least one neural network module, and the neural network module comprises a recurrent neural network and a gating recurrent unit network. According to the technical scheme, the problem that the protein expression effect is insufficient in an existing codon optimization technology is solved, codon optimization can be conducted on the protein sequence based on the model formed by the recurrent neural network and the gated circulation unit network, and the protein expression effect is improved.

The invention discloses a recombinant engineering bacterium for expressing fetuB and application of the recombinant engineering bacterium, and relates to the technical field of genetic engineering. According to the recombinant engineering bacteria, pichia pastoris serves as host bacteria, after fetuB genes are introduced, efficient expression of the fetuB in the pichia pastoris is achieved by optimizing codons and improving fermentation conditions, and the expression quantities of CBS7435 under the shake-flask culture condition are 320 + / -25 mg / L, SMD116 under the shake-flask culture condition is 280 + / -30 mg / L, and SuperMan5 under the shake-flask culture condition is 580 + / -45 mg / L. After fermentation conditions are optimized, the culture yield of a fermentation tank can reach 27.35 g / L. The fetuin B disclosed by the invention is good in biological activity, high in expression, simple in operation process and suitable for large-scale industrial production.

Related to the field of prevention and treatment of respiratory diseases, and disclosed a method for preparing a preventive mRNA vaccine against human respiratory syncytial virus. In some embodiments, the mRNA vaccine comprises an antigen-encoding mRNA and lipid nanoparticles. In some embodiments, the mRNA sequence utilizes an improved 5'UTR / 3'UTR sequence and codon optimization methods for efficient translation of the target protein. In some embodiments, the mRNA vaccine expresses an RSV F protein that is used as an antigen, wherein the sequence of the RSV F protein is optimized to achieve stable RSV pre-F protein expression. After immunization, the mRNA vaccine induces high levels of RSV F protein-specific binding antibody titer and neutralizing antibody titer in mice.

The invention belongs to the technical field of recombinant proteins, particularly discloses recombinant human fibronectin, a preparation method and application of the recombinant human fibronectin in cosmetics, and aims to solve the technical problems that existing recombinant fibronectin is small in expression quantity and relatively low in activity. The recombinant human fibronectin disclosed by the invention is recombined after an active region of natural fibronectin is intercepted, the recombinant human fibronectin is introduced into a vector after codon optimization, escherichia coli is used as an expression host cell, and the recombinant human fibronectin disclosed by the invention is obtained after fermentation and purification. The recombinant human fibronectin disclosed by the invention is high in expression quantity and relatively good in biological activity, can promote cell adhesion, proliferation or migration, and has a wide application prospect in the field of cosmetics for skin care, beauty and the like.

The present disclosure relates to a codon optimization technique based on token classification. In a model training phase, a collected protein sequence is tokenized by using a single amino acid as a token, and a nucleic acid sequence encoding a protein and corresponding to the protein sequence is tokenized by using a single codon as a token; a plurality of different synonymous codons in the tokenized nucleic acid sequence encoding the protein and corresponding to a single amino acid in the tokenized protein sequence are sorted in a descending order of frequency of occurrence in an expression system of the protein, and a classification label is set, thereby training a natural language model. In a prediction phase of the nucleic acid sequence encoding the protein, a protein sequence to be subjected to codon optimization is tokenized. The tokenized protein sequence is input into the trained natural language model to predict a classification label of a synonymous codon corresponding to a single amino acid in the tokenized protein sequence. An optimized nucleic acid sequence is obtained on the basis of a mapping relationship between the synonymous codon and the classification label thereof.

The invention discloses an mRNA (messenger Ribonucleic Acid) vaccine of varicella-zoster virus as well as a preparation method and application of the mRNA vaccine. According to the mRNA vaccine provided by the invention, a gE antigen sequence is subjected to codon optimization; then selecting a proper promoter, 5 'UTR, 3' UTR and 3 '-terminal PolyA to combine into complete mRNA nucleic acid for coding the varicella-zoster virus, screening to obtain two groups of mRNA nucleic acid for coding the varicella-zoster virus with both expression level and stability, and adopting a novel cationic lipid imidazopyridyl lipid compound to prepare a vaccine liposome. The targeting property in drainage of lymph nodes is improved, and the humoral immunity and cellular immunity levels excited by the vaccine in animal bodies are remarkably improved.

The invention discloses a construction and identification method of an influenza B DNA vaccine, which comprises the following steps: step 1, downloading sequence information of an influenza B recommended strain B / Austria / 1359417 / 2021 in vaccine strain recommended components in 2022-2023, carrying out codon optimization on an HA gene of the strain, and then carrying out gene synthesis, and naming the gene as BHA; 2, highly conservative sequences of HA protein, M2e protein and NA protein are selected for series combination, gene synthesis is performed after codon optimization, and the gene is named as G3; 3, respectively extracting plasmid DNAs of the synthetic fragments for later use; 4, amplifying a target fragment by using a PCR technology so as to connect a carrier; and 5, carrying out a target gene and vector connection experiment. According to the present invention, the HA protein sequence of the DNA vaccine can be rapidly replaced by using the restriction enzyme cutting site, the preparation method is simple, the immunogenicity is good, the cross protection is good, the preparation cost is low, the preparation requirement of the large-scale influenza vaccine can be met, and the important scientific significance and the important social value are provided.

The invention provides alkali-resistant glycosyl transferase and a coding gene, a biological material, a preparation method and application thereof. The glycosyl transferase has a specific amino acid sequence and can be efficiently expressed in escherichia coli through a genetic engineering means. The preparation method comprises the steps of codon optimization, recombinant plasmid cloning, expression strain transformation, induced expression, purification and the like. The obtained glycosyltransferase has significant application in preparation of gastrodin by enzymatic catalysis of p-hydroxybenzyl alcohol, and enzymatic hydrolysis conditions such as substrate concentration, enzyme concentration, UDP-glucose concentration and temperature, and buffer solution type and pH value are optimized. Particularly, when a glycine-sodium hydroxide buffer solution is used at the pH value of 9.5, the enzymolysis effect is remarkable. According to the alkali-resistant glycosyl transferase as well as the preparation method and the application thereof, a powerful tool is provided for biosynthesis of natural products such as gastrodin, and the alkali-resistant glycosyl transferase has an important industrial application value. Meanwhile, the biological material disclosed by the invention provides a basis for production and application of alkali-resistant glycosyl transferase.

The invention belongs to the technical field of immunology, and particularly relates to an anti-TNF-alpha nano antibody as well as a preparation method and application thereof. The anti-TNF-alpha nano antibody sequence with high affinity is obtained through four rounds of optimized phage screening, and further, the soluble expression quantity of the nano antibody is improved by utilizing a pET26a (+)-BL21 expression system and combining a codon optimization technology. The nano antibody can be used for preparing a reagent for treating or diagnosing TNF-alpha related diseases, and has extremely high practical application value.

The invention discloses a new enzyme with acylsulfonate transferase activity or a mutant thereof and application thereof, and belongs to the field of enzyme engineering. On the basis of an NCBI (National Center of Biotechnology Information) database, a new enzyme sequence is mined through a bioinformatics means, a new enzyme is obtained through codon optimization, gene synthesis and cell expression, the activity of acylsulfonate transferase is measured, and a corresponding mutant is obtained by using a directed evolution method. Compared with reported mouse acyl sulfonate transferase (ASTIV), the enzyme sequence obtained in the invention has better catalytic activity in a regeneration system of PAPS, and provides a new choice for enzymatic synthesis of PAPS.

The invention belongs to the technical field of biology, particularly relates to a vomitoxin detoxification enzyme and application thereof, and discloses a DON detoxification enzyme gene aadh sequence derived from acidobacteria, a DON detoxification enzyme coded by the DON detoxification enzyme gene aadh, a preparation method of the DON detoxification enzyme and application of the DON detoxification enzyme in degradation of vomitoxin. The preparation method comprises the following steps: performing codon optimization and gene synthesis on a DON detoxifying enzyme gene aadh; constructing a prokaryotic expression vector of the DON detoxifying enzyme gene aadh; expressing the prokaryotic expression vector in a prokaryotic host cell; the DON detoxifying enzyme provided by the invention can degrade vomitoxin, and can remove toxins in mildewed cereal raw materials and feeds.

The invention discloses acyl sulfonic acid transferase and application thereof, and belongs to the field of enzyme engineering. On the basis of an NCBI (National Center of Biotechnology Information) database, a new enzyme sequence is mined through a bioinformatics means to obtain the acyl sulfonate transferase derived from Podarcismuris, a new enzyme is obtained through codon optimization, gene synthesis and cell expression, and the activity of the acyl sulfonate transferase is measured. Compared with reported mouse acyl sulfonate transferase, the acyl sulfonate transferase provided by the invention has better catalytic activity in a regeneration system of PAPS, and a new choice is provided for enzymatic synthesis of PAPS.

The invention relates to the technical field of plant genetic engineering, in particular to application of an algae NySAT2 gene in regulation and control of alfalfa crude protein and detergent fiber. According to the invention, a sequence after codon optimization of an NySAT2 gene coding region is obtained by using gene synthesis and codon optimization technologies, an algae NySAT2 gene overexpression vector is constructed, a target gene is introduced into an alfalfa genome by using an agrobacterium rhizogenes mediated genetic transformation method, and a transgenic plant is obtained by using a genetic transformation system. Molecular identification and phenotypic analysis show that on the premise of keeping normal growth vigor (plant height and biomass have no significant difference from those of a wild type), the crude protein content of a medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased compared with that of the wild type medicago sativa plant, and meanwhile, the contents of acid detergent fibers and neutral detergent fibers are remarkably reduced, so that the content of the crude protein in the medicago sativa plant with the overexpressed NySAT2 gene is remarkably increased. The forage quality and the feeding value of the medicago sativa are effectively improved.