406 results about "Prokaryotic expression" patented technology

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody 4F6 for HPV16 type E7 protein detection and application. The heavy chain variable region CDR sequences and the light chain variable region CDR sequences of the antibody are SEQ ID NO.1-3 and SEQ ID NO.4-6 respectively, and the antibody can be specifically combined with HPV16 type E7 protein without cross reaction. A double-antibody sandwich ELISA detection system constructed based on the antibody is combined with a biotin-avidin amplification technology, the sensitivity reaches 100 pg / mL, and the antibody is suitable for rapid detection of the HPV16 type E7 protein in a cervical exfoliated cell sample. The invention further provides the HPV16 type E7 recombinant protein which is obtained through prokaryotic expression and has immunocompetence, and the HPV16 type E7 recombinant protein is applied to antibody screening and detection. Compared with nucleic acid detection, the method has a lower false positive rate, can be used for early screening of cervical cancer and precancerous lesions, and has a good application prospect.

The invention discloses three pattern recognition protein Hc beta GRP genes for preventing and treating fall webworms, dsRNA and application thereof. Specifically, the nucleotide sequences of the three pattern recognition protein Hc beta GRP genes for preventing and treating fall webworms are respectively SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3. The recombinant Hc beta GRP protein is successfully obtained through a prokaryotic expression system, an in-vitro bacteriostatic test proves that the recombinant Hc beta GRP protein, the Hc beta GRP protein and the dsRNA have direct bacteriostatic activity, dsRNA is designed based on a pattern recognition protein Hc beta GRP gene, a hyphantria cunea Hc beta GRP dsRNA interference system implemented by using an injection method is established, an obvious gene silencing effect is obtained by injecting the Hc beta GRP dsRNA interference system into hyphantria cunea, and the Hc beta GRP dsRNA interference system is applied to the hyphantria cunea. The technical support is provided for the application of the Hc beta GRP RNAi technology. Besides, the dsRNA of the hyphantria cunea Hc beta GRP is combined with the biocontrol bacteria by utilizing an established hyphantria cunea Hc beta GRP RNA interference system, so that the control effect of the biocontrol bacteria can be remarkably improved.

The invention discloses L-arabinose isomerase, engineering bacteria and application, the amino acid sequence of the L-arabinose isomerase is shown as SEQ ID NO: 3, and the gene sequence for coding the L-arabinose isomerase is shown as SEQ ID NO: 2. On the basis of a genome of a Peribacillussp.S4 strain, a gene for coding the L-arabinose isomerase is excavated, and a stable prokaryotic expression system is constructed by performing codon optimization on the gene, so that the large-scale controllable production of the L-arabinose isomerase is realized, the yield of the enzyme is remarkably increased, the production cost of the enzyme is reduced, and the method is suitable for industrial production. A stable and economical enzyme source is provided for the production of D-tagatose; the L-arabinose isomerase expressed by the constructed engineering bacteria can significantly improve the yield of D-tagatose, and industrialization of D-tagatose is facilitated.

The invention relates to the technical field of biological medicines, in particular to a recombinant rabies virus nano-antibody fused with a solubilizing oligopeptide and a cell penetrating peptide and application of the recombinant rabies virus nano-antibody in preparation of rabies prevention and treatment products. The recombinant rabies virus nano-antibody fused with the solubilizing oligopeptide and the cell penetrating peptide comprises a humanized rabies virus nano-antibody, and the C end of the humanized rabies virus nano-antibody is fused with a solubilizing tag peptide P17 and the cell penetrating peptide. The recombinant rabies virus nano antibody utilizes a prokaryotic expression system, is low in cost, has excellent cell membrane and BBB penetrating ability, can enter a central nervous system from veins for virus neutralization treatment, and has an efficient passive immune effect.

The invention discloses methanogen lyase homologous with a PeiR lyase sequence and application of the methanogen lyase, and aims to solve the problem that PeiR lyase protein of a methanogen cell wall peptide bond can participate in a biological process of hydrolyzing archaea cell walls, effectively kill methanogen and reduce methane production, and protein homologous with the PeiR lyase protein has potential methane reduction potential. According to the invention, a series of biological information software is utilized to identify protein sequences coded by methanogens virus from rumen microbiome sequencing data, and sequence homology between the proteins and PeiR protein is further analyzed to obtain a series of methanogens lyases. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through the detection of crude enzyme liquid, the effectiveness of the lyase in the aspect of reducing the methane yield is verified in an in-vitro gas production test.

The invention discloses a pertussis recombinant protein and a monoclonal antibody and application thereof.The pertussis recombinant protein is synthesized by selecting an S1 subunit of pertussis PT and a part of sequences of FHA protein, the pertussis recombinant protein is expressed through a prokaryotic expression system and purified, the protein serves as an antigen to immunize a mouse, and the pertussis recombinant protein is obtained. Positive hybridoma cell strains 3B10, 3D9 and 5F7 capable of stably secreting the antibody are obtained by adopting a hybridoma technology, the preservation numbers are respectively CCTCC NO: C202567, CCTCC NO: C202568 and CCTCC NO: C202569, and the obtained monoclonal antibody has relatively high affinity and detection sensitivity to pertussis PT and FHA recombinant proteins, is used for detecting a bordetella pertussis antigen, is strong in specificity and high in sensitivity, can be used for instant detection, and can be applied to clinical application. And the method has a relatively high application value.

The invention discloses methanogen lyase homologous with high-dimensional characteristics of PeiW lyase and application of the methanogen lyase, and aims to solve the problem that PeiW lyase protein of methanogen cell wall peptide bonds can participate in the biological process of hydrolyzing archaea cell walls, effectively kill methanogen and reduce methane production, and protein homologous with the protein has potential methane reduction potential. According to the method, a series of bioinformatics software is utilized, a protein sequence coded by methanogens virus is identified from a large amount of rumen microorganism virus group sequencing data, the high-dimensional homology of a deep learning model between the protein sequence and PeiW is analyzed, and two methanogens lyases similar to the high-dimensional characteristics obtained by the PeiW model are obtained. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through detection of a crude enzyme solution, the effectiveness of the lyase in the aspect of reducing the yield of methane is verified in an in-vitro gas production test.

The invention discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody generated by the herbicide-resistant protein GAT hybridoma cell strain and an application of the herbicide-resistant protein GAT hybridoma cell strain. The herbicide-resistant protein GAT hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46333. The preparation method comprises the following steps: a) purifying prokaryotic expression to obtain GAT recombinant protein; b) immunizing animals: immunizing BALB / c mice by taking the GAT recombinant protein as an antigen; c) cell fusion: splenocytes of the immune BALB / c mice are collected and fused with SP2 / 0 cells; and d) cell strain establishment: carrying out subcloning through a limited dilution method, carrying out ELISA detection after 5-7 days of subcloning until a hybridoma cell strain capable of stably secreting a positive antibody is screened out, and carrying out amplification, re-culture and storage. The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for detection of herbicide-resistant protein GAT in transgenic crops, so that development, seed production and seed conservation of transgenic gat crops are supported, and stable industrial promotion of the transgenic gat crops and sustainable application of related transformants are guaranteed.

The invention discloses methanogen lyase with the same protein family annotation as PeiR lyase and application of the methanogen lyase, and the methanogen lyase is obtained through homologous screening of Pfam protein family database annotation based on known PeiR lyase characteristics aiming at methanogen cell wall peptide bonds. The PeiR lyase protein can participate in the biological process of hydrolyzing archaea cell walls, methanogens are effectively killed, methane production is reduced, and the protein homologous with the protein has potential methane reduction potential. According to the invention, by integrating metagenome sequencing data of rumen microorganisms, screening methanogenic bacterium virus protein by using series bioinformatics software, and combining Pfam functional domain annotation to carry out homologous relationship analysis, a series of lyase is finally identified and is successfully expressed in a prokaryotic expression system, and meanwhile, an in-vitro gas production experiment result also shows that the methanogenic bacterium virus protein can be successfully expressed in a prokaryotic expression system. The crude enzyme liquid can significantly reduce the methane generation amount.

The invention discloses methanogen lyase homologous with high-dimensional characteristics of PeiR lyase and application of the methanogen lyase and aims to solve the problems that PeiR lyase protein of methanogen cell wall peptide bonds can participate in the biological process of hydrolyzing archaea cell walls, methanogen is effectively killed, methane production is reduced, and protein homologous with the protein has potential methane reduction potential. According to the invention, a series of biological information software is utilized to identify protein sequences coded by methanogen viruses from a large amount of rumen microbiome sequencing data, and high-dimensional homology between the proteins and PeiR proteins based on a deep learning model TM-Vec is further analyzed, so that a series of methanogen lyases are obtained. In order to realize the expression of the lyase, multiple segments of primers are designed to synthesize a target sequence, and the target sequence is successfully expressed in a prokaryotic expression system. Through the detection of crude enzyme liquid, the effectiveness of the lyase in the aspect of reducing the methane yield is verified in an in-vitro gas production test.

The invention relates to the technical field of biology, in particular to a preparation method and application of a recombinant human long-acting interleukin-22 binding protein. The amino acid sequence of the recombinant human long-acting interleukin-22 binding protein is as shown in SEQ ID No. 1. According to the method for preparing the IL-22BP recombinant protein with high biological activity by utilizing an Escherichia coli prokaryotic expression system, a novel fusion protein is designed, a sequence for coding human IL-22BP is connected with a sequence for coding ABD to construct an IL-22BP-ABD fusion gene, the IL-22BP-ABD fusion gene is induced to be expressed in Escherichia coli, inclusion body protein is separated, and the IL-22BP recombinant protein with high biological activity is obtained. The target protein is obtained through denaturation, purification and renaturation, and the biological activity of the IL-22BP-ABD is verified through in-vivo and in-vitro experiments.

The invention discloses an influenza C virus HEF protein polyclonal antibody preparation method, which comprises: carrying out signal peptide prediction, antigen epitope screening and hydrophobicity analysis through bioinformatics, constructing a recombinant prokaryotic expression vector pET32a-HEF by using a seamless cloning technology, and efficiently expressing the HEF protein in Escherichia coli to obtain the influenza C virus HEF protein polyclonal antibody. The inclusion body protein yield is improved by optimizing conditions; and purifying by nickel column affinity chromatography to obtain high-purity recombinant protein, emulsifying the high-purity recombinant protein and a Freund's adjuvant, immunizing a Japanese white rabbit, immunizing for three times, and collecting high-titer serum. The method breaks through detection limitation caused by antigen tag concealment in a eukaryotic expression system, the obtained polyclonal antibody is high in specificity, the eukaryotic expression HEF protein can be accurately recognized, and a key tool is provided for ICV virus-like particle identification, vaccine research and development and diagnosis technologies. Compared with a traditional method, the scheme is simple and convenient to operate, low in cost and high in antibody titer, and has remarkable application value.

The invention discloses cloning and functional identification of a liverwort flavone synthetase gene, and belongs to the technical field of gene engineering. According to the invention, one flavone synthase I (flavones synthase I, FNS I) is obtained through screening, and is named as MpFNS I; a prokaryotic expression system is utilized, an MpFNS I recombinant protein is obtained through purification, and the result of in-vitro enzyme activity identification shows that an MpFNS I enzyme substrate is relatively wide in selectivity, has a flavone synthetase function and also has a part of flavonol synthetase function. The liverwort flavonoid has a significant difference on affinity and catalytic efficiency of different substrates, the in-vivo function of the liverwort flavonoid is verified by utilizing heterologous expression of arabidopsis thaliana, experimental evidence is provided for comprehensively analyzing a biosynthetic pathway of the liverwort flavonoid, and meanwhile, a research target is provided for deeply researching a mechanism for synthesizing and responding to UV-B radiation of the liverwort flavonoid.

The invention discloses a method for realizing high-efficiency selenium labeling of protein, which comprises the following steps of: in the process of expressing plasmids containing target protein through a prokaryote expression system, realizing high-efficiency replacement of cysteine sulfur atoms in the target protein by selenium atoms by regulating and controlling the ratio of selenium to sulfur elements in a culture medium; on the premise of keeping a good growth state of cells, the selenium labeling efficiency can reach 86%, and the protein expression quantity is stabilized at about 1mg / g (protein / thallus wet weight); structural analysis shows that the secondary structure of the obtained selenium-labeled protein is basically consistent with that of natural protein and is not obviously changed. The method is easy and convenient to operate, high in labeling efficiency, high in protein yield, good in repeatability and suitable for structural biology research such as nuclear magnetic resonance, an effective means is provided for research of the functional mechanism of sulfur atoms in biomacromolecules (including small peptides, polypeptides, peptide fragments, proteins and protein or peptide complexes) with selenium as a probe, and the method has a wide application prospect. Good scientific research application prospects and industrial popularization values are realized.

The invention relates to the technical field of immunodetection, and discloses a bovine nodular skin disease virus chimeric antigen and application thereof in an ELISA (enzyme-linked immuno sorbent assay) detection kit. According to the chimeric antigen, a gene of a dominant region of a bovine nodular skin disease virus antigen is selected to design a chimeric antigen gene, meanwhile, sequence optimization is performed, a chemical synthesis method is used for synthesizing a coding gene of the chimeric antigen of the bovine nodular skin disease virus, and then the chimeric antigen is obtained in a prokaryotic expression mode. An ELISA antibody detection kit developed on the basis of the chimeric antigen has extremely high sensitivity and specificity, can rapidly and specifically detect bovine nodular skin disease virus antibodies in serum, is suitable for monitoring and checking the nodular skin diseases of cattle in large-scale breeding, and has relatively high practical value.

The invention belongs to the technical field of plant disease control, and particularly relates to preparation and application of diaphorina citri sourced antibacterial peptide DcAMP1. The amino acid sequence of the antibacterial peptide DcAMP1 is SEQ ID NO. 1, and the nucleotide sequence of the gene for coding the antibacterial peptide DcAMP1 is SEQ ID NO. 2. The prokaryotic expression plasmid is adopted to construct the recombinant plasmid containing the gene encoding the antibacterial peptide DcAMP1, prokaryotic expression of the antibacterial peptide DcAMP1 is achieved, and the method is simple and easy to implement. After being purified, the antibacterial peptide DcAMP1 obtained through prokaryotic expression has an obvious antibacterial effect on a liberobacter asiaticum related bacterium sinorhizobium fredii, and the titer of the liberobacter asiaticum in a sweet orange tree with the liberobacter asiaticum can be obviously reduced.

The invention discloses a polyclonal antibody of a grouper iridovirus SGIV VP101 gene and application of the polyclonal antibody. The polyclonal antibody is prepared from an SGIVVP101 gene, and the nucleotide sequence of the VP101 gene of an optimized codon is as shown in SEQ ID NO. 1. The polyclonal antibody of the SGIV VP101 gene recombinant expression protein prepared by the invention has good specificity, not only can specifically recognize a prokaryotic expression product of the SGIV VP101, but also can be used for quantitative and positioning analysis of the SGIV VP101 in vivo and in vitro. The successful preparation of the polyclonal antibody of the recombinant expression protein of the SGIV VP101 gene lays a foundation for the establishment of an SGIV immunological detection method, the quantitative and positioning analysis of the protein of the VP101 gene and the analysis of gene functions.

The invention discloses an application of an AP2-F2 protein polyclonal antibody as a cryptosporidium female gamete positioning tag antibody. The AP2-F2 protein of cryptosporidium parvum is subjected to truncated prokaryotic expression, protein is purified to immunize an ICR mouse, a polyclonal antibody is prepared, ELISA and WB detection of antibody titer are carried out to prove that the polyclonal antibody is effective, then indirect immunofluorescence detection of the antibody is carried out to determine that the polyclonal antibody is located on the female gametes of cryptosporidium parvum, and the polyclonal antibody can be used as a label to locate the female gametes of cryptosporidium parvum. The method plays an important role in localization research of unknown protein of cryptosporidium.

The invention belongs to the technical field of plant disease control, and particularly relates to preparation and application of diaphorina citri sourced antibacterial peptide DcAMP2. The amino acid sequence of the antibacterial peptide DcAMP2 is SEQ ID NO. 1, and the nucleotide sequence of the gene for coding the antibacterial peptide DcAMP2 is SEQ ID NO. 2. The prokaryotic expression plasmid is adopted to construct the recombinant plasmid containing the gene encoding the antibacterial peptide DcAMP2, prokaryotic expression of the antibacterial peptide DcAMP2 is achieved, and the method is simple and easy to implement. After being purified, the antibacterial peptide DcAMP2 obtained through prokaryotic expression has an obvious antibacterial effect on a liberobacter asiaticum related bacterium sinorhizobium fredii, and the titer of the liberobacter asiaticum in a sweet orange tree with the liberobacter asiaticum can be obviously reduced.

The invention discloses a human growth hormone detection antibody and a preparation method thereof. The amino acid sequence of the detection antibody is one of SEQ ID NO. 1 and SEQ ID NO. 2; the preparation method comprises the following steps: extracting RNA (Ribonucleic Acid), performing reverse transcription, amplifying obtained cDN A, inserting into escherichia coli, and constructing a human growth hormone primary library; the M13KO7 auxiliary bacteriophage invades the human growth hormone primary library to construct a bacteriophage primary library; carrying out three rounds of solid phase panning on the phage primary library, and carrying out monoclonal screening, ELISA detection and sequencing to obtain a target gene sequence; and synthesizing the target gene sequence into a prokaryotic expression plasmid, introducing the prokaryotic expression plasmid into escherichia coli, and adding IPTG for expression to obtain the detection antibody. The detection antibody has the characteristics of low economy, strong stability and simple subsequent operation, and solves the problem that animal immunization is always needed to prepare the antibody in the subsequent process in human growth hormone detection.

The invention belongs to the field of biology, and particularly relates to a rhododendron molle heat shock protein RmHSP20-4 gene and application thereof. The RmHSP20-4 participates in response of rhododendron molle to high-temperature stress, and can be used for screening high-temperature tolerance germplasm of rhododendron molle and evaluating high-temperature tolerance of sheep. The gene is transferred into a prokaryotic expression strain, the growth of the transgenic strain under the stress of CdCl2 and high temperature is obviously improved, and the RmHSP20-4 gene and a protein coded by the RmHSP20-4 gene provide candidate genes for the cultivation of Rhododendron molle heavy metal resistance, high temperature stress resistance and other related varieties. A new candidate gene resource is provided for cultivating transgenic plants or microorganisms with excellent shapes such as heavy metal resistance and high temperature stress resistance, and an important reference is also provided for mining heat-resistant genes of azaleaceae plants.

The invention discloses a sheep paratuberculosis specific detection kit and application, and relates to the field of biological detection.The kit comprises an elisa plate coated with a recombinant Mycobacterium paratuberculosis membrane protein antigen, the nucleotide sequence of the recombinant Mycobacterium paratuberculosis membrane protein antigen is shown as SEQ ID NO: 1, and the amino acid sequence of the recombinant Mycobacterium paratuberculosis membrane protein antigen is shown as SEQ ID NO: 2; a confining liquid; a serum diluent; carrying out enzyme labeling on a second antibody; a TMB color developing solution and a stop solution; according to the sheep paratuberculosis specific detection kit disclosed by the invention, the recombinant Mycobacterium paratuberculosis membrane protein antigen is adopted as a core detection component, and the antigen has high specificity and can accurately identify paratuberculosis positive serum and effectively distinguish positive and negative samples; meanwhile, prokaryotic expression and purification are carried out on the specific membrane protein of the mycobacterium paratuberculosis for the first time, and a novel sheep paratuberculosis indirect ELISA diagnosis method with high sensitivity and specificity based on the protein as an antigen is established.

The application discloses an elagatis bipinnulatus IGF2 gene and application thereof, wherein the nucleotide sequence of the elagatis bipinnulatus IGF2 gene is shown as SEQ ID NO:1, the amino acid sequence of the protein coded by the IGF2 gene is shown as SEQ ID NO:2. A prokaryotic expression vector containing the gene and a corresponding recombinant strain are successfully constructed, a recombinant IGF2 protein is prepared through a high-efficiency expression system, and the recombinant IGF2 protein is applied to preparation of a product for regulating fish feeding, preparation of an additive with the function of promoting fish feeding and preparation of artificial feed with the function of promoting fish feeding. The recombinant IGF2 protein can significantly enhance the expression of feeding-related genes of the elagatis bipinnulatus and significantly improve the feeding amount.

The invention discloses a paralichthys olivaceus rhabdovirus G protein tandem antigen epitope peptide and application thereof, and belongs to the field of fish molecular immunology. The amino acid sequence of the tandem antigen epitope peptide is shown as SEQ ID NO: 1. The preparation method comprises the following steps: (1) firstly, analyzing structural characteristics of HIRRV-G protein, predicting B cell antigen epitopes of the HIRRV-G protein, screening the antigen epitopes with advantages on the basis of a prediction result, and synthesizing the antigen epitopes; (2) screening a candidate peptide fragment with high affinity through an enzyme-linked immunosorbent assay; and (3) sequentially connecting the high-affinity peptide fragment sequences meeting the requirements by using a GPGPG connexon, cloning the connected sequences into a pET-28a prokaryotic expression vector, and performing induced expression to obtain the tandem antigen epitope peptide. Compared with a full-length G protein, the tandem antigen epitope peptide is smaller in molecular weight, higher in stability and higher in hydrophilicity; a large number of specific antibodies can be induced in fish bodies, and the death rate of the paralichthys olivaceus infected by viruses is remarkably reduced. The method can be used for HIRRV diagnosis detection reagent and subunit vaccine development.

The invention discloses a purification method for removing endotoxin in prokaryotic expression recombinant protein, which belongs to the technical field of bioengineering, and comprises the following steps: adding 0.1% Triton X-114 in an ultrasonication stage, carrying out ice-bath ultrasonication, after the recombinant protein is combined with a Ni-NTA affinity column, washing with 50 column volumes of a combination buffer solution containing 0.1% Triton X-114, and carrying out elution by using an elution buffer solution containing 0.1% Triton X-114 in the presence of an elution buffer solution containing 0.1% Triton X-114 in the presence of an elution buffer solution containing 0.1% Triton X-114 in the presence of an elution buffer solution containing 0.1% Triton X-114; and washing with a 4-DEG C binding buffer solution with 20 column volumes, and subsequently eluting the target protein with imidazole with different gradients. Then, regulating the pH value of the purified recombinant protein solution to 2-4 or more than 10 through 100mM hydrochloric acid or sodium hydroxide, dissociating and decomposing endotoxin, carrying out oscillation treatment at room temperature for 2-4 hours, then removing the endotoxin in the recombinant protein through ultrafiltration, retaining an endotube protein solution, and detecting the content of protein and the endotoxin in the recombinant protein; the method is easy and convenient to operate and low in cost, the endotoxin content in the recombinant protein prepared through prokaryotic expression can be reduced by 400 times on the premise that immune recognition of the recombinant protein and serum is not affected, and the phenomenon that strong cross reaction occurs between immune serum and escherichia coli due to high endotoxin residues in the recombinant protein is reduced.

The invention discloses a recombinant 2.1 d subtype swine fever E2 protein, a subunit vaccine and application of the subunit vaccine, and belongs to the technical field of genetic engineering. According to the 2.1 d subtype swine fever E2 protein, 344-375 amino acids of the 2.1 d subtype swine fever E2 protein are removed, and 1-343 amino acids of the 2.1 d subtype swine fever E2 protein are reserved, so that the most important antigenic epitope with a protective effect of the 2.1 d subtype swine fever E2 protein can be reserved, the space structure of the recombinant 2.1 d subtype swine fever E2 protein is basically not influenced, and the immunogenicity of the 2.1 d subtype swine fever E2 protein can be reserved to the greatest extent; in addition, the recombinant 2.1 d subtype swine fever E2 protein can be efficiently expressed in a prokaryotic expression system; furthermore, the soluble expression level of the recombinant 2.1 d subtype swine fever E2 protein can be remarkably improved through co-expression of the molecular chaperone, and after the recombinant 2.1 d subtype swine fever E2 protein is prepared into a subunit vaccine, the subunit vaccine has a good protection effect on 2.1 d subtype swine fever virus infection.

The invention belongs to the technical field of biological detection, and particularly relates to a preparation method of lipopolysaccharide-beta-1, 3-glucan binding protein and application of the lipopolysaccharide-beta-1, 3-glucan binding protein in preparation of a broad-spectrum colloidal gold detection product for dermatophytes. The prokaryotic expression system induces the procambarus clarkii-sourced lipopolysaccharide-beta-1, 3-glucan binding protein to be efficiently expressed in an inclusion body form, the optimized inclusion body purification-renaturation process is combined to realize batch preparation, and the colloidal gold chromatography technology is integrated, so that the procambarus clarkii-sourced lipopolysaccharide-beta-1, 3-glucan binding protein is obtained. The bottlenecks of the traditional detection method in sensitivity, broad spectrum and field applicability are broken through.

The invention relates to the technical field of biology, in particular to an anti-porcine CD82 protein monoclonal antibody, a hybridoma cell strain and application, the monoclonal antibody is prepared by the following steps: expressing and purifying a CD82 (EC2) region by utilizing a prokaryotic expression system, preparing by taking the purified recombinant protein as an immunogen, and carrying out indirect ELISA (enzyme-linked immuno sorbent assay) and Flow Cytometry screening to obtain the monoclonal cell strain capable of secreting an IgG2a type antibody. The purified antibody is detected through an indirect ELISA method, sensitive reaction still exists when the antibody concentration reaches 31.25 ng / ml, PAM cells capable of expressing endogenous CD82 protein are detected through flow cytometry, and the result shows that the mAb has good marking capacity for the PAM cells, indicating that the mAb of the CD82 protein capable of being used for flow cytometry is successfully prepared. A biological tool is provided for further analyzing the function and mechanism of the CD82 protein.

The invention belongs to the field of gene engineering, and provides a mycoplasma ovipneumoniae multi-epitope fusion protein and preparation and application thereof.The inventor firstly successfully predicts antigen epitopes of three virulence genes and performs tandem expression on the predicated antigen epitopes to prepare a fusion protein antigen, and the fusion protein antigen is used for preparing the mycoplasma ovipneumoniae multi-epitope fusion protein. A secondary structure, a tertiary structure, physicochemical properties (including hydrophilicity, stability, PI value and the like), antigenicity, sensitization and the like of the fusion protein are predicted through biological online software, and the success rate of a test is increased; meanwhile, the fusion protein connected in series with the three virulence genes of the mycoplasma ovipneumoniae is prepared into a subunit vaccine, and a good immune protection effect is also achieved. The recombinant mycoplasma ovipneumoniae multi-epitope fusion protein prokaryotic expression plasmid pET28a < + >-EHP is successfully constructed, tandem expression of mycoplasma ovipneumoniae on an escherichia coli prokaryotic system is achieved, and an expression product can obtain a good immune protection effect.

The invention provides a subunit vaccine composition for porcine epidemic diarrhea, porcine delta coronal and porcine rotavirus as well as a preparation method and application of the subunit vaccine composition, and is characterized in that the subunit vaccine composition comprises prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein, viral subunit protein and pharmaceutically acceptable adjuvants; wherein the amino acid sequence of the prokaryotically expressed swine erysipelas filamentous bacillus SpaA protein is as shown in SEQ No.1, and the viral subunit protein is selected from one or more of porcine epidemic diarrhea virus S protein, porcine delta coronavirus S protein and porcine rotavirus VP8 protein. The vaccine composition disclosed by the invention has the advantages of strong immunogenicity, good safety, no immune interference, high neutralizing antibody titer and long antibody duration.