149 results about "Codon optimization" patented technology

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 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 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.

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 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 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.

The invention discloses a method for synthesizing alpha-arbutin through biotransformation, and belongs to the technical field of biochemistry. A gene cloning technology is utilized, sucrose phosphorylase Spase in leuconostoc mesenteroides and sucrose phosphorylase gtfa in bifidobacterium adolescentis are screened and found through a Deep Molecules and DLKcat algorithm on a Uniprot database, novopro is utilized for codon optimization, then the sucrose phosphorylase Spase and the sucrose phosphorylase gtfa are expressed on a plasmid vector PET-28a and are converted into an escherichia coli BL21 (DE3) strain, and the sucrose phosphorylase Spase and the sucrose phosphorylase gtfa in the bifidobacterium adolescentis are obtained. An engineering strain for heterologous expression of the gene is constructed by taking the gene as a chassis cell. According to the process, sucrose and hydroquinone are taken as co-substrates, and bioconversion is carried out under the action of sucrose phosphorylase from different sources to obtain alpha-arbutin.

The invention discloses a green algae expression method and application of recombinant human serum albumin, and belongs to the technical field of biology. Aiming at the problems of high cost and incomplete post-translational modification of the existing recombinant protein expression system, the invention provides a technical scheme for efficiently expressing human serum albumin by using green algae. The method comprises the following steps: carrying out codon optimization on a human serum albumin gene sequence, introducing a recombinant vector into green algae cells, and screening transgenic algae strains through a culture medium containing a resistance agent, culturing the transgenic algal strain, splitting the algal cells and separating the recombinant albumin. According to the method, efficient production of the human serum albumin is achieved through a green alga expression system, and the product has a correct folding structure and biological activity and is mainly used for preparing medical plasma substitutes, drug carriers and biological agents.

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

The invention discloses a bivalent subunit vaccine for preventing infection of haemophilus parasuis and porcine circovirus type 2. A core antigen combination of the vaccine comprises at least one antigen protein from haemophilus parasuis, and the antigen protein is selected from Ferrin, OppA and Hem-SAP and is combined with a porcine circovirus type 2 Cap protein. Wherein the NCBI (National Center of Biotechnology Information) login number of the Ferrin, the NCBI login number of the OppA and the NCBI login number of the Hem-SAP are WP160414389.1, ACL32731.1 and WP035493594.1 respectively. The antigen protein is subjected to codon optimization, is expressed and purified through a prokaryotic expression system, and is emulsified with ISA201 or a Freund's adjuvant to prepare the vaccine. Animal experiments prove that the vaccine can excite high-level antigen specificity IgG, cell factors IFN-gamma and white IL-4 in an immune animal body, namely, specific Th1 and Th2 type immune responses are generated. The vaccine can generate an immune protection rate of up to 80% when attacked by a serum type 5 haemophilus parasuis virulent strain, and generates an effective antibody response to the porcine circovirus type 2. The vaccine provided by the invention has the advantages of definite components, good safety and strong immune protection force.

The invention belongs to the technical field of biology, and provides an application of alpha-amylase protein derived from nephroplasmosis in culture of nephroplasmosis, and the alpha-amylase protein is obtained by synthesizing a gene sequence of alpha-amylase as shown in SEQ ID NO.2 after codon optimization and then transfecting the gene sequence to an expression vector. And carrying out bacterial transformation, culture, induction and crushing. The invention also provides a method for inducing the formation of the reproductive cyst of the reniform worm. It is found for the first time that alpha-amylase protein specifically induces reproductive cysts to be formed, the biomass of the nephroplasmosis is increased, the growth generation of the nephroplasmosis is shortened, and the method is suitable for species such as distended nephroplasmosis and nephroplasmosis henslouianum and fits natural ecological scenes; complex equipment is not needed, the cost is low, large-scale production is easy, the stability is high, and the method can be widely applied to soil ecological restoration, agricultural microbial resource development and basic research related to nephroplasmosis.

This invention belongs to the fields of molecular biology and bioinformatics, specifically relating to a truncated bovine virus-encoded protein phage library, its construction method, and applications. This library covers all 162 known bovine viruses that host bovine subfamily viruses, containing 51,794 non-redundant peptide coding sequences of 56 amino acids each. These peptides are fused and displayed on the surface of the T7 phage capsid, with all coding sequences having a uniform total length of 200 bp. The library construction was achieved through sequence acquisition, truncation and redundancy removal, codon optimization and DNA synthesis, and in vitro ligation and packaging with T7 phage. The resulting library exhibits a positivity rate of 93% and a titer of 2.8 × 10⁻⁶. 10 PFU / mL. This invention features an optimized single-round high-throughput screening process adapted to clinical bovine serum samples, enabling efficient identification of bovine virus B-cell epitopes and providing tools and candidate targets for bovine virus vaccine development and diagnostic reagent development.

The invention belongs to the technical field of bioengineering, and particularly relates to a kluyveromyces marxianus engineering strain for recombinant expression of hydroxylated human III-type collagen and application of the kluyveromyces marxianus engineering strain. The Kluyveromyces marxianus engineering strain of the human type III collagen is obtained by carrying out recombinant expression construction on proline hydroxylase and a human type III collagen gene in the Kluyveromyces marxianus; according to the invention, two truncation designs and codon optimization are carried out on the existing natural human III type collagen amino acid sequence to obtain the human III type collagen C3S1 and C3S2, and the recombinant secretory expression in the kluyveromyces marxianus engineering strain is realized. Like hydroxylation modification of the human III type collagen, the human III type collagen C3S1 and the human III type collagen C3S2 which are subjected to recombinant expression have proline hydroxylation characteristics. The recombinant human III-type collagen is prepared by using the kluyveromyces marxianus engineering strain, the method is simple, the production efficiency is high, and large-scale production is easy.

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

The application discloses application of PgUGT29 in synthesis of kudou saponin D3. Specifically, the application includes cloning, codon optimization, protein expression and purification of the gene PgUGT29 and application of the enzyme activity of the protein. The nucleotide sequence of the kudou PgUGT gene is shown as SEQ ID NO. 1, the optimized nucleotide sequence is shown as SEQ ID NO. 2, and the encoded amino acid sequence is shown as SEQ ID NO. 3. The kudou PgUGT29 gene in the application has the function of specifically catalyzing kudou saponin D to generate kudou saponin D3, and can be used for generating kudou saponin D3, improving the quality of kudou and improving the pharmacological activity of kudou.

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

The invention discloses an application of a transcription factor ChZF7 of a basophilic microalgae Chlorella sp. BLD in regulation and control of salt tolerance of microalgae. According to the invention, the coding gene of the transcription factor ChZF7 is transferred into chlamydomonas reinhardtii by using a transgenic technology after intron insertion and codon optimization, so that transgenic chlamydomonas reinhardtii is obtained. The salt tolerance of the chlamydomonas reinhardtii with the over-expressed ChZF7 gene is remarkably improved, the accumulation of neutral fat and active oxygen under the stress of 225mM salt is less than that of the chlamydomonas reinhardtii in a control group, the content of penetrating substance proline is increased, in addition, the change of cell components in the chlamydomonas reinhardtii under the stress of salt can be weakened by the over-expressed ChZF7 gene, and the salt tolerance of the chlamydomonas reinhardtii is improved. The transcription factor ChZF7 and the gene coded by the transcription factor ChZF7 can regulate and control the salt tolerance of the chlamydomonas reinhardtii, a foundation is laid for genetic improvement and directed molecular breeding of the microalgae, the transcription factor ChZF7 plays an important role in improvement of the salt tolerance of the microalgae, reutilization of salinized water resources is facilitated, and the transcription factor ChZF7 and the gene coded by the transcription factor ChZF7 have wide application prospects.

The invention relates to the technical field of biosynthesis, in particular to a biosynthesis process of adenosine triphosphate and equipment thereof. Comprising the following steps: optimizing an adenosine triphosphate synthetase gene sequence, constructing an expression vector, and introducing the expression vector into a yeast host cell; sub-packaging the engineering yeast group in a pore plate for parallel culture, detecting the acid yield and substrate conversion rate data of each micropore, and screening out elite strains; fermenting elite strains, and dynamically feeding materials by an online monitoring and feedback control system; the yeast strain is subjected to precise transformation through codon optimization and gene overexpression driven by a strong promoter; elite strains are quickly identified by adopting high-throughput screening in cooperation with miniaturized parallel fermentation and real-time optical detection; in the fermentation process, a self-control feedback fed-batch technology is introduced, and a culture environment is stabilized in an optimal physiological state through real-time monitoring and dynamic regulation, so that the production potential of engineering bacteria is fully exerted, and the yield of adenosine triphosphate, the substrate conversion rate and the product purity are improved.

The invention discloses a recombinant collagen, a coding gene and a preparation method and application thereof, and belongs to the technical field of gene engineering and protein engineering. On the basis of an amino acid sequence of human VII type collagen, the amino acid sequence is designed by means of bioinformatics and the like, a new recombinant collagen is constructed, further, a pichia pastoris strain for efficiently expressing the recombinant collagen is constructed by optimizing a gene sequence through codons, and the recombinant collagen is obtained. The recombinant collagen has good biocompatibility and biological activity, can remarkably promote proliferation, adhesion and migration of L929 cells, and has wide application prospects in multiple fields such as food, medicines, cosmetics or instrument products.

The invention provides an expression vector, an expression system and a preparation method of a recombinant larosidase protein, and belongs to the technical field of recombinant protein preparation, the recombinant expression vector of the larosidase protein provided by the invention comprises an initial vector and a codon-optimized human-derived larosidase coding gene; the nucleotide sequence of the human-derived larosidase coding gene is as shown in SEQ ID NO. 1; according to the recombinant expression system, Chinese hamster ovary cells are taken as host cells, and the recombinant expression vector is transfected. According to the invention, a gene sequence more suitable for expression of Chinese hamster ovary cells is obtained through codon optimization, and then transposon-mediated stable integration is carried out; high-density suspension culture and downstream purification processes are utilized to realize efficient and stable expression of the recombinant larosidase in CHO cells, and the recombinant larosidase with high yield, high activity and high purity can be obtained.

This invention relates to the field of biochemical technology, specifically to a recombinant batroxobin, its preparation method, and its applications. The invention relates to a recombinant batroxobin that uses a short sequence of α-mating factor signal peptide and a short sequence of Bat as repeating units, and connects multiple repeating units through short sequences of tandem IRES elements. The tandem IRES element short sequences are selected from one or more of the following amino acid sequences: amino acid sequences expressed by any of the sequences shown in SEQ ID No. 1-4 and SEQ ID No. 21. The synergistic effect of IRES, stable sequences, and codon optimization overcomes multiple rate-limiting steps from gene integration to protein synthesis, resulting in a significant increase in the recombinant expression level of batroxobin.

The invention relates to a methanophilic genetically engineered bacterium for producing gamma-polyglutamic acid (gamma-PGA) as well as a construction method and application of the methanophilic genetically engineered bacterium, EQU2420670 gene is knocked out on a host bacterium genome, and meanwhile, an exogenous gamma-PGA synthetase encoding gene cluster pgsBCAE under the control of an inducible arabinose promoter (Pbad) is introduced at the site. The construction method comprises the following steps: obtaining a multi-cistron structure DNA fragment pgsBCAE for coding gamma-PGA synthetase through artificial gene synthesis in combination with a codon optimization strategy, and constructing the expression module Pbad-pgsBCAE under the control of a Pbad promoter. A suicide vector pK18mobsacB carrying a SacB sucrose reverse selection system is used for constructing a gene replacement vector, upstream and downstream homologous arms of an EQU2420670 gene on a genome of methanophilic bacteria M. buryatase 5GB1 are designed on two sides, an EQU2420670 site is replaced by a Pbad-pgsBCAE expression module, the methanophilic genetically engineered bacterium PGA is obtained, and the methanophilic genetically engineered bacterium PGA is applied to production of gamma-PGA and has important economic and social significance; the gamma-PGA produced by the engineering methanophilic bacteria by using methane as a carbon source and an energy source can be used as a cosmetic additive; the invention provides a novel low-carbon biosynthesis approach for commercial production and application of gamma-PGA, and has a good industrial application prospect.

The invention discloses a pearl gentian grouper IgM monoclonal antibody, which is characterized in that screened pearl gentian grouper hybridoma No. 41 is sequenced to obtain nucleotide and amino acid sequences of variable regions of a heavy chain and a light chain of the targeted pearl gentian grouper IgM monoclonal antibody respectively shown as SEQ ID NO.1-12, the heavy chain subtype of the monoclonal antibody is IgG type, the light chain is kappa type, and the light chain subtype of the monoclonal antibody is kappa type. Through codon optimization, recombinant pATX1 vectors of heavy-chain and light-chain antibodies are respectively constructed with a constant region sequence of a murine antibody. The Epinephelus lanceolatus IgM monoclonal antibody is expressed through co-transfection XtenCHO cell in-vitro culture, the chimeric immune globulin is good in reaction characteristic and high in titer (512000 or above) and is easier to store compared with a hybridoma cell mode, and the risk of losing the specific antibody is avoided.

The invention discloses a novel codon optimization method based on precise transcriptome sequencing and analysis thereof, and belongs to the technical field of bioinformatics. In order to improve the expression quantity of a target gene, corresponding accurate transcriptome data is obtained according to the development period of transgenic expression and parts such as tissues and organs, and efficient expression genes are screened to serve as a host genome expression library; the expression abundance of the gene is comprehensively considered when the use frequency of the codon of the host genome is determined, and the gene optimized by the novel codon optimization method increases the stability of the insecticidal protein gene RNA and improves the expression level of the protein. The invention provides better reference for gene modification and transformation, and provides a novel codon optimization method for high-efficiency expression of transgenes in plants.

The invention discloses a mycobacterium tuberculosis mRNA (messenger Ribonucleic Acid) vaccine as well as a construction method and application thereof, and belongs to the technical field of biological medicines. The technical problem to be solved is to provide a novel mycobacterium tuberculosis mRNA vaccine which is low in dosage, high in safety and lasting in immune memory. According to the technical scheme, the preparation method is characterized by comprising the following steps: carrying out tandem expression on antigens for coding Hsp65 and Mpt83, constructing mRNA molecules by combining tPA signal peptide (SEQ ID NO: 1), an MITD sequence (SEQ ID NO: 5) and flexible Linker (SEQ ID NO: 6-7), and carrying out codon optimization (an mRNA secondary structure and GC content) and a lipid nanoparticle (LNP) encapsulation process to finally obtain the novel mycobacterium tuberculosis mRNA vaccine.

The present invention discloses a laccase, its gene and application, belonging to the technical field of genetic engineering. The laccase gene of the present invention is obtained by codon optimization of the laccase gene of Cerrena unicolor, and the nucleotide sequence is as shown in SEQ ID NO.2. The laccase gene provided by the present invention is recombinantly expressed by transforming Aspergillus niger, and high-activity laccase can be mass-produced industrially. The highest enzyme activity in shake flask culture reaches 39 U / ml, and after culturing in a 7L fermenter for 125 h, the total laccase activity reaches 9234 U / ml. At the same time, the laccase provided by the present invention and the mediator methyl syringate have a significant destructive effect on zearalenone, which is beneficial to grain transportation and storage.

The present application relates to a kind of monoclonal antibody that can identify human Topo II alpha antigen, secretes cell strain, its preparation method and its use in immunodetection.The above technical solution selects the amino acid of 1301-1531 of Topo II alpha protein as antigen peptide, codon optimization becomes the gene fragment suitable for expression in escherichia coli BL21, and finally the recombinant protein includes Topo II alpha protein fragment and histidine protein label.The recombinant protein is immunized to mouse, is fused by cell, screening and subcloning, and obtains mouse hybridoma cell strain, and the anti Topo II alpha protein monoclonal antibody secreted by the cell strain is obtained.The antibody obtained by the present application has high specificity, sensitivity, and can specifically recognize the cell expressing Topo II alpha protein, and is suitable for immunological detection, especially immunohistochemical detection.