9699results about "Genetic engineering" patented technology

The invention belongs to the technical field of biology, and relates to an escherichia coli engineering strain for producing L-isoleucine through a methyl malic acid way as well as a construction method and application of the escherichia coli engineering strain. According to the invention, Escherichia coli BW25113 is used as a chassis strain, and a methyl malic acid synthase high-activity mutant gene cimA3.7, an isopropyl malic acid isomerase gene GsleuCD, a 3-isopropyl malic acid dehydrogenase gene AfleuB and a leucine dehydrogenase gene Lsleudh are over-expressed; a methyl malic acid absorption protein gene is over-expressed, and a branched chain amino acid transport system is modified to improve the exosome of the L-isoleucine; all methyl malic acid pathway related genes are integrated to a genome of a chassis strain, and a plasmid-free L-isoleucine production strain is constructed; the copy number of a key gene on a genome is optimized, and the yield of the engineering strain L-isoleucine is increased by introducing a non-oxidative glycolysis approach. The yield of the L-isoleucine after the engineering strain is fermented for 34 hours in a fermentation tank is 56.6 g / L.

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 a biosynthesis system for high-yield 1, 6-hexamethylenediamine and application of the biosynthesis system, and belongs to the technical field of biology. According to the method, key genes which are excavated and optimized in a 1, 6-hexamethylenediamine biosynthetic pathway are co-expressed in a single cell module or a multi-cell module, and different enzyme elements expressed by the cell module are subjected to enzyme cascade reaction, so that 1, 6-hexamethylenediamine can be efficiently synthesized by taking caprolactam or 6-aminocaproic acid as a substrate; furthermore, after the carboxylic acid reductase MaCAR is mutated, the catalytic activity of the carboxylic acid reductase MaCAR can be remarkably improved, and the yield of the 1, 6-hexamethylenediamine is further improved; in addition, the biosynthesis system disclosed by the invention directly takes the cell module as a whole-cell catalyst, and an enzyme purification step is avoided, so that the biosynthesis system has the advantage of low cost.

The present invention belongs to the field of biological detection technology, specifically relating to a monoclonal antibody combination for detecting HPV18 E7 protein and its application. This combination consists of monoclonal antibodies 5F8 and 6G6, wherein the heavy and light chain variable regions of 5F8 and 6G6, respectively, contain specific complementarity-determining region sequences (as shown in SEQ ID NOs. 1 to 12), conferring high specificity for HPV18 E7 protein. Verification using a double-antibody sandwich ELISA and a biotin-avidin amplification system demonstrated that this antibody combination specifically recognizes HPV18 E7 protein, exhibits no cross-reactivity with other high-risk HPV oncoproteins, and exhibits a detection sensitivity of 1 ng / mL. This combination can be used to construct detection systems such as ELISA kits and immunochromatographic test strips, suitable for rapid screening of HPV18 E7 protein in cervical exfoliated cell samples.

The invention provides a recombinant strain for controllably synthesizing high-proportion 4-hydroxybutyric acid (4HB) polyhydroxyalkanoate (PHA) as well as a construction method and application of the recombinant strain. The recombinant strain disclosed by the invention can be used for efficiently producing high-proportion poly (3-hydroxybutyrate-4-hydroxybutyric acid) (P34HB for short) by using glucose as a unique carbon source or a 4HB structure related or non-related carbon source in amplification production of a shake flask, a 7L fermentation tank, a 100L fermentation tank and a 5000L fermentation tank.

The invention belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody for specific recognition of type II grass carp reovirus VP4 protein, an eukaryotic expression method and application. According to the invention, a target gene S6 is transfected to an HEK293 cell for expression and purification, a New Zealand white rabbit is immunized in cooperation with a Freund's adjuvant, and the rabbit-derived recombinant monoclonal antibody capable of specifically recognizing the GCRV-II VP4 protein is obtained by using ELISA, single B cell screening and eukaryotic expression technologies. The antibody has strong specificity, provides support for further establishment of a specific II-type grass carp reovirus diagnosis technology, and has great application value in development of GCRV-II related scientific research.

The invention belongs to the technical field of biological medicines, and particularly relates to a composition for treating male erectile dysfunction and application thereof. The oyster polypeptide is prepared by enzymolysis of oyster protein, can effectively promote proliferation of testicular interstitial cells, enhance and improve sexual desire and improve sexual behavior, has certain clinical application value, has the advantages of simple preparation, easy absorption and the like, and is easy to popularize and apply.

The invention provides recombinant Escherichia coli. The recombinant Escherichia coli comprises the following multiple modifications: knocking out genes such as iclR, lpxM, ldhA, nadR, ptsP and the like; the expression of genes such as narJ, setA and the like is enhanced. The Escherichia coli disclosed by the invention can rapidly intake a glucose raw material, and can rapidly grow under various culture conditions. Therefore, the method has potential application value in the field of fermentation.

The invention discloses application of a TaLBD30 protein and a coding gene thereof in regulating and controlling a wheat plant type. The invention belongs to the technical field of biology, and particularly relates to application of TaLBD30 protein and a coding gene thereof to regulation and control of wheat plant types. The protein is any one of the following proteins: A1) a protein with an amino acid sequence as shown in SEQ ID No: 1; a2) a protein which is obtained by substitution and / or deletion and / or addition of amino acid residues on the protein of A1), has 80% or more of identity with the protein of A1) and has the same function as the protein of A1); a3) a fusion protein obtained by connecting a protein tag to the N terminal or / and the C terminal of A1) or A2). TaLBD30 is a transcription factor with transcription inhibition activity, and compared with a wild type, the TaLBD30 gene overexpression shows that after TaLBD30 overexpression, the wheat plant height and ear length are remarkably reduced.

The invention belongs to the technical field of nano antibodies, and particularly relates to a nano antibody specifically bound with AAV9 and application of the nano antibody. The nano antibody provided by the invention can be specifically combined with the AAV9 adeno-associated virus without being combined with other AAV serotypes such as AAV2, AAV5 and AAV8, so that high-sensitivity and high-specificity detection of the AAV9 adeno-associated virus is realized. In practical application, the nano-antibody coupled solid-phase carrier provided by the invention is adopted as an affinity filler, the AAV9 adeno-associated virus can be effectively captured and mildly eluted, the virus purification and recovery effects are effectively improved, and the nano-antibody coupled solid-phase carrier is suitable for high-purity recovery of the AAV9 adeno-associated virus.

The invention discloses a gene and a protein for adjusting tobacco polyphenol aroma substances and application of the gene and the protein. The gene is a tobacco NtMYB28 gene, and the CDS sequence of the tobacco NtMYB28 gene is as shown in SEQ ID No. 1. The invention further discloses application of the tobacco NtMYB28 gene in regulating the content of endogenous polyphenol substances in tobacco and regulating and controlling the metabolic pathway of phenylpropane in tobacco. The key gene NtMYB28 capable of simply and effectively adjusting the content of polyphenol substances in tobacco and the metabolic pathway of phenylpropane is found, and the gene can be applied to efficient cultivation of new high-aroma tobacco materials and development and utilization of new high-value functional tobacco germplasm.

The invention relates to the technical field of biological detection, in particular to a monoclonal antibody combination for HPV18 type E6 protein detection and application. The provided combination is composed of 5G3 and 2C7, and the amino acid sequences of complementary determining regions of variable regions of a heavy chain and a light chain of the combination are clear and are respectively shown as SEQ ID NO.1-12. The antibody combination has high specificity and sensitivity, the lowest detection limit can reach 100 pg / ml, and cross reaction with other HPV subtypes is avoided. According to a double-antibody sandwich ELISA and biotin-avidin amplification detection system constructed based on the combination, the signal intensity and the detection accuracy are remarkably improved, and the combination is suitable for rapid detection of the HPV18 type E6 protein in a cervical exfoliated cell sample and has application value in early diagnosis of cervical cancer, risk stratification, vaccine research and development and curative effect evaluation.

The invention discloses a monoclonal antibody BY017 combined with African swine fever virus P30 protein and application of the monoclonal antibody BY017. The invention provides an IgG antibody which is composed of a light chain and a heavy chain. CDR1, CDR2 and CDR3 in the heavy chain variable region are sequentially shown as the 47 to 54 , the 72 to 79 and the 118 to 126 in SEQ ID NO: 1; cDR1, CDR2 and CDR3 in a light chain variable region are sequentially as shown in the 49 to 60 , the 78 to 80 and the 117 to 124 in SEQ ID NO: 3. The invention also protects the application of any one of the IgG antibodies in preparation of products. The product has the functions that the product is used for being combined with an African swine fever virus P30 protein fragment; the antibody is used for binding African swine fever virus P30 protein; the antibody is used for binding African swine fever virus. The IgG antibody provided by the invention can be used for multiple purposes of western blot, immunohistochemistry, flow cytometry, ELISA (Enzyme-Linked Immunosorbent Assay) and the like, and can be used as an important tool for ASFV detection and structural analysis of P30 protein of ASFV detection.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-human carbohydrate antigen CA125 monoclonal antibody and application thereof. The amino acid sequences of CDR1-3 on a light chain variable region of the monoclonal antibody are respectively shown as SEQ ID NO.3-5, and the amino acid sequences of CDR1-3 on a heavy chain variable region of the monoclonal antibody are respectively shown as SEQ ID NO.8-10. The monoclonal antibody provided by the invention can specifically recognize and bind to a natural carbohydrate antigen CA125 expressed by human cells or tissues, has good affinity and relatively high sensitivity of binding of the antibody and the antigen, has strong anti-interference ability to complex non-target protein components in the cells / tissues, is beneficial to significantly improving the accuracy and reliability of immunodetection of human CA125 protein, and has good application prospects. The method is suitable for high-specificity, high-sensitivity and high-reliability detection of the human CA125 protein, and has good applicability in a plurality of detection systems such as immunoblotting, immunoprecipitation, immunohistochemistry and the like.

The invention discloses a self-replicating RNA (Ribonucleic Acid) molecule, a porcine rotavirus RNA vaccine and application thereof, and relates to the technical field of vaccines. According to the present invention, the coding sequence of the porcine rotavirus G9P23 type VP4 protein is optimized, such that the fact that the saRNA prepared by using the nucleotide sequence represented by the SEQ ID NO: 10 can secrete the high porcine rotavirus G9P23 type VP4 protein antigen after the cell transfection is found; after the optimized porcine rotavirus G9P23 type VP4 protein is used for immunizing experimental animals, the level of neutralizing antibodies generated by the animals can be improved, and the challenge protection effect is enhanced. Therefore, the nucleic acid molecule provided by the invention can be used for developing drugs and vaccines for causing protective immune response in experimental animal bodies. Compared with a traditional vaccine, the vaccine provided by the invention has good immunogenicity and safety, and has a good application prospect.

The invention provides a group of specific antibodies for detecting botulinum toxin A and application thereof, and belongs to the technical field of botulinum toxin detection. The invention provides a group of specific antibodies for detecting botulinum toxin A or antigen binding fragments thereof. The specific antibodies or the antigen binding fragments thereof comprise a rabbit monoclonal antibody 1F11 or an antigen binding fragment thereof and a rabbit monoclonal antibody 2A10 or an antigen binding fragment thereof. The time-resolved immunodetection kit prepared by the invention has good detection sensitivity, specificity, repeatability and sample matrix tolerance, the detection range is 0.02 ng / mL-10 ng / mL, the detection limit is 0.02 ng / mL, cross reaction with other serotype botulinum toxins (B, E and F) does not exist, and the time-resolved immunodetection kit is not influenced by sample matrixes such as ham sausages and thick broad-bean sauce. The detection system established by the invention has a good application prospect in detection of botulinum toxin type A.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to an SCRE10 gene for improving rice disease resistance and application of the SCRE10 gene, the base sequence of the SCRE10 gene is shown as SEQ ID NO.1, and the amino acid sequence of the SCRE10 gene is shown as SEQ ID NO.2. Through construction of a dexamethasone induced expression SCRE10 transgenic rice plant, it is found that the transgenic rice can significantly induce PR gene expression and active oxygen outbreak, and the resistance of rice to false smut and bacterial leaf blight can be improved. The invention proves that heterologous inducible expression of the Ustilaginoidea virens SCRE10 gene has the function of positively regulating the disease resistance of the rice, and the SCRE10 gene can be used for improving the disease resistance of the rice, which is of great significance to the creation of disease-resistant germplasm of the rice.

The invention discloses a broad-spectrum monoclonal antibody for resisting porcine epidemic diarrhea virus S protein and application of the broad-spectrum monoclonal antibody. Specifically, firstly, two monoclonal antibodies GC33D8-1 and GC33A4-3 which have broad-spectrum binding characteristics with the porcine epidemic diarrhea virus S protein are obtained, the GC33D8-1 antibody has good universality and can have strong affinity with different strains, and the GC33A4-3 antibody can be bound for 3 times. Secondly, the two monoclonal antibodies are respectively used as a capture antibody and a detection antibody, a double-antibody sandwich ELISA method is established, and the double-antibody sandwich ELISA method is applied to PEDV vaccine S protein quantification, has better parallelism compared with other antibodies, and can be widely applied to detection of different strains. The method can be used for detecting semi-finished products and finished products, can be used for process establishment and optimization of porcine epidemic diarrhea virus vaccine antigens, vaccine quality evaluation and stability monitoring, has the advantages of high sensitivity, high accuracy, high repeatability and high flux, and solves the problems that broad-spectrum detection cannot be carried out at present and an S protein ELISA method only can be used for qualitative judgment.

This invention belongs to the field of biological detection technology, specifically relating to a monoclonal antibody combination for detecting feline NT-proBNP protein and its applications. The monoclonal antibody combination of the present invention comprises monoclonal antibodies 3A6 and 5H5. These two antibodies possess unique heavy and light chain variable region complementarity-determining regions, respectively, and their amino acid sequences have been fully defined (SEQ ID NOs. 1-12). This antibody combination can specifically recognize and efficiently bind to feline NT-proBNP protein, making it suitable for the development of a highly sensitive and specific diagnostic tool for feline cardiomyopathy. By constructing a double-antibody sandwich ELISA assay system, the minimum detection limit can reach 100 pg / mL, and it exhibits excellent specificity and stability.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody combination for detecting measles virus nucleoprotein and application of the monoclonal antibody combination. The combination is composed of monoclonal antibodies 5E2 and 5G9 and can efficiently and specifically recognize measles virus nucleoprotein, amino acid sequences of complementary determining regions (CDR) of heavy chain and light chain variable regions of the combination are clear and are shown as SEQ ID NO.1-12 respectively, high specificity and high affinity of the antibodies are guaranteed, the antibody combination can effectively capture and detect target antigens, and the antibody combination can be used for detecting measles virus nucleoprotein. The risk of non-specific binding and cross reaction is obviously reduced. The combination shows excellent sensitivity and specificity in detection platforms such as colloidal gold immunochromatography, and provides a stable and reliable biological recognition tool for rapid and accurate detection of measles viruses.

The invention relates to the technical field of antibodies, and provides an anti-CD45 antibody for detecting a white blood cell common antigen based on flow cytometry and application of the anti-CD45 antibody, the antibody comprises at least one of the following CDRs: a heavy chain variable region CDR: an amino acid sequence of SEQ ID NO: 1-3 or a conservatively modified form thereof; and a light chain variable region CDR: SEQ ID NO: 4-6 or an amino acid sequence in a conservative modification form thereof. The antibody or the antigen binding fragment thereof can efficiently and specifically recognize CD45, and can effectively detect the CD45.

This invention provides a humanized monoclonal antibody that binds to advanced glycation end product (AGE) modified proteins or peptides on a cellular surface. [Solution] A humanized monoclonal antibody for advanced glycation end products, comprising at least one amino acid sequence selected from a group consisting of eight specific amino acid sequences, each with a different sequence. The antibody may be bound to a carboxymethyllysine-modified protein or peptide on a cell. The antibody can be used in a cell separation process, for example, in magnetic cell separation.

There is provided a fluorinase variant thereof having at least 70% sequence identity to a sequence MSDLGX6TDDSVAQCKGLMLSICPX24VX26IX28DX30CHX33MTPX37DVVEGARYIVDLPR X52FPEGTVFATTTYPATGTX70X71RSVAX76RX78KX80AALGGARGQX90AGSGX95GX97E RAEGX103YIYIAPNNGLLTX116VIEEHGYX124EAYEVSX131TX133VIPX137X138PEPTFYSR EMVAIPSAHLAAGFPLX163X164VGRX168LX170DX172EIVRFEX179X180KX182X183X184VX186G X188X189LX191GX193X194X195X196X197DHPFGNX204WTNX208HRTDLEKAGIX219YX221TX223X 224KX226VX228DGVLX233FX235LPLX239PTFADAX246X247X248GX250PVX253YX255NSRGYLX2 62X263ARNAAX269LAYPYNLX277AGX280SVX283VTX286A (SEQ ID NO: 1), wherein X is a natural amino acid, and wherein the variant comprises one or more mutations at a site selected from the group consisting of an ion-egress site, a S-Adenosyl-L-Methionine (SAM) binding site, an ion-binding site (IBS), and a conserved site. Also disclosed are polynucleotides encoding the variants, vectors comprising the polynucleotides encoding the variants, and host cells comprising the vectors thereof. Also disclosed are Methods for producing the variants, methods of catalyzing the fluorination of a compound, uses of the fluorinase variants, and methods of treatment using the fluorinase variants thereof.

Antibodies and antibody fragments that bind to polyfluoroalkyl and perfluoroalkyl species (PFAS) are described for use in biosensors for detecting PFAS compounds in environmental samples. A detector comprising the biosensor and a kit for use with the detector are also described.

The invention provides an antibody for resisting CD19 on the surface of a porcine B cell or an antigen binding fragment thereof as well as a composition and application thereof. Specifically, the invention provides an anti-porcine B cell surface CD19 antibody or an antigen binding fragment thereof, which can effectively bind to CD19 on the porcine B cell surface. The invention also provides an antibody drug conjugate containing the antibody or the antigen binding fragment thereof, and the antibody drug conjugate can be applied to screening of porcine B cells and marking of CD19 antigens on the surfaces of the porcine B cells, and has a good application prospect.

Provided herein are systems, compositions, and methods for generating immunogenic peptides or epitopes from tumor associated antigens (e.g., in vivo or ex vivo). Polynucleotides (e.g., genes) encoding the tumor associated antigens may be edited at selected target sites by nucleobase editors comprising a catalytically-inactive Cas9 and a cytosine deaminase, leading to the expression of heteroclitic or cryptic peptides that are more immunogenic than the native peptide derived from the tumor associated antigens. The heteroclitic or cryptic peptide elicit strong tumor-specific immune response (e.g., T-cell response or B-cell response), which inhibits tumor growth and metastasis.

The invention belongs to the technical field of biological detection, and relates to an antibody composition aiming at pTau217 protein detection and application thereof, and the antibody composition aiming at pTau217 protein detection comprises a target pTau217 protein antibody and a target Tau protein C-terminal antibody. Meanwhile, based on the antibody composition aiming at pTau217 protein detection, the invention further provides a kit for early diagnosis of the Alzheimer's disease. The invention provides an antibody composition aiming at pTau217 protein detection, which is stronger in detection specificity and higher in sensitivity, and an application of the antibody composition.

The invention discloses a specific antibody MR-34 for targeting a coronavirus MERS-CoV spike protein RBD (Radial Basis Disease). The sequences of the CDRs of the antibody disclosed by the invention are as shown in SEQ ID NO. 1 to SEQ ID NO. 6. The antibody provided by the invention contains CDRs with a specific sequence, and can be specifically combined with an MERS-CoV spike protein RBD structural domain. The antibody provided by the invention is strong in targeting property and high in binding activity, and the antibody prepared by the preparation method provided by the invention is high in purity and can be prepared on a large scale. On the whole, the antibody disclosed by the invention can be used for developing and optimizing an MERS-CoV infection detection method.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody 1D3 for detecting a porcine epidemic diarrhea virus antibody and application of the monoclonal antibody 1D3. The CDR sequences of the heavy chain variable region are respectively shown as SEQ ID NO.1-3, and the CDR sequences of the light chain variable region are respectively shown as SEQ ID NO.4-6. The antibody can specifically recognize PEDV / S1 protein, binding of the antibody can be competitively blocked by a PEDV specific neutralizing antibody in serum, and the antibody is suitable for establishing a blocking ELISA method. A detection tool based on the antibody has high specificity and sensitivity, and the blocking rate to PEDV positive serum is higher than 50%. The invention can be used for preparing a detection kit or evaluating the immune effect of a vaccine, and provides an efficient and accurate technical means for PEDV infection diagnosis and immune monitoring.