12955results about "Genetic engineering" patented technology

The invention provides an anti-human serum albumin nano antibody and application thereof, and belongs to the technical field of antibodies. The invention provides an anti-human serum albumin nano antibody HSA-32 with relatively high affinity, and the anti-human serum albumin nano antibody HSA-32 is subjected to humanized transformation, so that three obtained humanized anti-human serum albumin nano antibodies can be used for reducing the immunogenicity of the HSA-32 and improving the immunogenicity of the HSA-32; and high specificity and high affinity of combination with the human serum albumin and stability and solubility of the antibody are maintained. The anti-human serum albumin nano antibody can be used for separation, purification and detection of human serum albumin and separation and purification of polypeptide, protein or antibody drugs fused with the anti-human serum albumin nano antibody. Meanwhile, the anti-human serum albumin nano-antibody can effectively prolong the half-life period of polypeptide, protein and antibody drugs fused with the anti-human serum albumin nano-antibody, and can be used for developing long-acting polypeptide, protein or antibody drugs.

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 immunology, and particularly relates to an anti-IL-6 nano antibody, a recombinant nano antibody, a recombinant vector and application. The anti-IL-6 nano antibody disclosed by the invention comprises a heavy chain variable region containing CDR1, CDR2 and CDR3, and has the functions of identifying and combining IL-6. The invention also discloses a recombinant nano antibody containing the nano antibody, a recombinant vector and a detection agent, and applications of the nano antibody, the recombinant nano antibody, the recombinant vector and the detection agent in preparation of drugs for treating IL-6 related diseases. The nano antibody, the recombinant nano antibody and the recombinant vector provided by the invention can recognize and combine IL-6, and have a potential effect of treating IL-6-related diseases.

The invention belongs to the technical field of immunology, and particularly relates to a targeted IL-6 biological nano antibody and application thereof. The invention discloses a nano antibody for targeting IL-6, which comprises a heavy chain variable region containing CDR1, CDR2 and CDR3, and has the functions of identifying and combining IL-6. The invention also discloses a composition containing the nano antibody or the IL-6 binding molecule, and an application of the composition containing the nano antibody and the IL-6 binding molecule in preparation of drugs for treating IL-6 related diseases. The nano antibody, the BCMA binding molecule and the composition can recognize and bind IL-6, and have a potential effect of treating IL-6 related diseases.

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 invention discloses an application of a TaNLP-3B gene and a related biological material thereof in regulating and controlling the protein content of plant seeds. The gene TaNLP-3B is cloned from wheat, the gene TaNLP-3B is overexpressed in wheat Fielder to obtain a TaNLP-3B transgenic overexpressed plant, and a detection result shows that the protein content in wheat seeds can be remarkably increased by more than 40% through TaNLP-3B gene overexpression, so that the TaNLP-3B gene has an effect of regulating and controlling the protein content in the plant seeds, and the protein content in the wheat seeds can be increased by more than 40%. The method can be applied to plant breeding or quality improvement.

The invention discloses an acyltransferase mutant and application of the acyltransferase mutant in synthesis of N-acetyl-trans-4-hydroxyproline, and belongs to the technical field of enzyme engineering. The acyltransferase mutant provided by the invention contains 10th and 95th mutations on the basis of a corresponding amino acid sequence, such as a sequence as shown in SEQ ID NO.1, or contains 62nd or 64th mutations while containing the 10th and 95th mutations. Compared with an original enzyme before mutation, the acyltransferase mutant obtained by the invention has higher acyltransferase activity, and the conversion rate of trans-4-hydroxy-L-proline is greatly improved, so that the yield of N-acetyl-trans-4-hydroxy proline is further improved, and the yield of N-acetyl-trans-4-hydroxy proline is improved. After the surfactant and the metal ions are added, the yield of the N-acetyl-trans-4-hydroxyproline synthesized by the mutant D62K / D10V / D95I under the catalysis is up to 18.7 g / L.

The invention discloses development, preparation and application of a carbapenemase immunochromatography kit, and belongs to the technical field of biological medicines. According to comparison of amino acid sequences of five common carbapenemase variants of KPC, IMP, NDM, VIM and OXA48 types, the immunogen is prepared from conserved epitope sequences obtained respectively. A mouse is immunized, and a specific antibody pair which is strong in affinity and can be combined with multiple variants is obtained through multi-platform screening. On the basis, the carbapenemase quintuplet detection kit is developed by utilizing an immunochromatography method, has the advantages of high sensitivity, good specificity, capability of covering various drug-resistant variants, capability of detecting multiple targets at one time and rapidness and convenience in detection, and provides important help for clinical auxiliary diagnosis and medication guidance.

The present invention relates to a recombinant yeast with high squalene production, a construction method and a use thereof, wherein the recombinant yeast is a saccharomyces cerevisiae that expresses or overexpresses HMGR gene, ADA gene, LEU2 gene, MVA pathway gene and squalene synthase gene ERG9. The present invention integrates acetaldehyde dehydrogenase gene ADA and HMG-CoA reductase gene tHMGR in multiple copies in saccharomyces cerevisiae, integrates the screened high-activity phosphoketolase gene PK and phosphotransacetylase gene PTA, weakens epoxysqualene synthesis, strengthens the mevalonate pathway, overexpresses yeast endogenous squalene synthase gene, etc. to obtain a recombinant yeast with high squalene production, so that the production in the shake flask stage of the production process exceeds 5g / L, and the production in the fed batch fermentation stage exceeds 50g / L, breaking through the current bottleneck of microbial synthesis of squalene production, and is suitable for practical promotion and application.

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

The invention discloses preparation of a targeting LYPD3 antibody and application of the targeting LYPD3 antibody in tumor immunotherapy. The antibody or antigen binding fragment thereof comprises a heavy chain variable region having CDR-H1, CDR-H2 and CDR-H3 and a light chain variable region having CDR-L1, CDR-L2 and CDR-L3 wherein CDR-H1, CDR-H2 and CDR-H3 comprise the amino acid sequences of SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5, respectively, and CDR-L1, CDR-L2 and CDR-L3 comprise the amino acid sequences of SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively.

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 belongs to the technical field of gene engineering, and particularly relates to application of a ZmDIR11 gene in regulating and controlling salt stress resistance of corn. After salt stress treatment, it is found that the ZmDIR11 gene mutant corn plant growth vigor and material accumulation are significantly lower than those of a wild type B73 material, an oxidative metabolism regulation path, proline accumulation, chlorophyll synthesis, lignan accumulation and a Na < + > / K < + > transport pathway are inhibited, and related metabolic indexes are significantly lower than those of the wild type B73 material. It is shown that the ZmDIR11 gene positively regulates and controls the salt-resistant metabolic pathway of the corn in the corn, and the salt tolerance of the corn is reduced after the ZmDIR11 gene is mutated. The research provides a new gene resource for corn salt-tolerant breeding by utilizing gene engineering.

The invention relates to a biotechnology, and particularly discloses a plasmin degradation product D-dimer antibody and application thereof. According to the application, a single B cell antibody preparation technology and a flow cytometry sorting method are applied to the development of a blood coagulation in-vitro diagnosis fibrinolysis system detection antibody for the first time, and a mammalian cell expression system (293F) is adopted to successfully express and obtain three strains of D-dimer monoclonal antibodies (antibody numbers are DD-1, DD-2 and DD-3) with biological activity. Compared with the on-sale Roch D-dimer antibody, the three antibodies have the same affinity with the DD antigen, and can identify and detect the D-dimer in plasma with high specificity and high sensitivity; meanwhile, the probe has no cross reaction with fibrinogen, primary fibrinolytic substances (X, Y and D) and soluble fibrin (SF), and can be used for qualitative or quantitative detection of D-dimer.

The invention discloses an anti-human hepatitis B e antigen monoclonal antibody and application thereof, and relates to the technical field of hepatitis B detection. In a heavy chain variable region of the monoclonal antibody, an amino acid sequence of a complementarity determining region CDR1-3 is as shown in SEQ ID NO.1-3 or has at least 95% of homology with the sequence as shown in SEQ ID NO.1-3; in the light chain variable region, amino acid sequences of complementary determining regions CDR1 and CDR3 are shown as SEQ ID NO.4 and SEQ ID NO.5 or have at least 95% of homology with the sequences shown as SEQ ID NO.4 and SEQ ID NO.5, and an amino acid sequence of a complementary determining region CDR2 is FAS. The monoclonal antibody provided by the invention can well recognize a natural hepatitis B e antigen, and the titer of a supernatant cultured by a B cell can reach 1: 10000, so that the monoclonal antibody can be completely applied to detection research of the human hepatitis B e antigen. The chemiluminescence reagent prepared by using the monoclonal antibody has the advantages of high specificity, strong anti-interference capability, high detection sensitivity, good stability and the like, almost has no leak detection, and can replace an imported reagent.

The invention relates to the technical field of biomedicine, in particular to an anti-influenza A virus NP protein antibody pair and application thereof. The antibody pair comprises two monoclonal antibodies; the monoclonal antibody comprises a heavy chain variable region and a light chain variable region; one monoclonal antibody in the monoclonal antibody pair is named as a 10G10 antibody, and the other monoclonal antibody is named as a 11F8 antibody. The invention also discloses a nucleic acid molecule for coding the monoclonal antibody and a hybridoma cell strain for secreting the monoclonal antibody. In addition, the invention also discloses a kit for detecting influenza virus protein, which comprises the monoclonal antibody pair, the 10G10 antibody is used as a capture antibody, and the 11F8 antibody is used as a labeled antibody. The antibody for resisting the influenza A virus NP protein provided by the invention has relatively good specificity on the influenza virus protein.

Provided are an apolipoprotein E detection reagent and the use thereof. The reagent comprises at least one of an ApoE2 protein detection reagent, an ApoE3 protein detection reagent and an ApoE4 protein detection reagent. One of a capture antibody and a detection antibody used for detecting ApoE2 protein or ApoE4 protein is a specific monoclonal antibody, and the capture antibody and the detection antibody used for detecting ApoE3 are both specific monoclonal antibodies. Genotyping of ApoE and quantitative detection of different genotypes of proteins are performed on the basis of an immunological detection method. The detection method has the advantages of being simple, easy to operate, short in time and cheap. A genotype detection result is highly consistent with a fluorescence PCR result. Genotyping of ApoE (6 types) can be performed, which is used to replace the nucleic acid detection and guide medication, and clarify the correlation of the ApoE4 genotype homozygosity / heterozygosity and the protein concentration of ApoE4 with AD.

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 relates to methods of modulating immune cells in a patient by altering microbiota of the patient. The invention also relates to methods of modulating treatments or therapies in a subject organism by altering microbiota of the subject. The invention also relates to cell populations, systems, arrays, cells, RNA, kits and other means for effecting this. In an example, advantageously selective targeting of a particular species in a human gut microbiota using guided nucleic acid modification is carried out to effect the alteration.

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 a hog cholera virus broad-spectrum neutralizing monoclonal antibody and application thereof, and belongs to the field of immune globulin. Specifically disclosed is an antibody or an antigen binding fragment thereof, comprising a heavy chain variable region and a light chain variable region; the amino acid sequences of CDR1, CDR2 and CDR3 in the heavy chain variable region sequentially comprise the 9th site to the 15th site, the 33rd site to the 40th site and the 79th site to the 90th site in SEQ ID No.1; the amino acid sequences of the CDR1, the CDR2 and the CDR3 in the light chain variable region sequentially comprise the 21 to 31 sites, the 49 to 51 sites and the 88 to 97 sites in SEQ ID No.3. The monoclonal antibody disclosed by the invention can be subjected to specific reaction with all genotype strains of the hog cholera virus, and the hog cholera virus E2 antibody can be detected by utilizing a blocking ELISA method established by the monoclonal antibody. The monoclonal antibody provided by the invention can be used for CSFV antigen identification and serological antibody detection kit development, and provides antibody resources for accurate diagnosis, prevention and control and purification of CSFV.

The invention provides a Ustilaginoidea virens gene UvASE2 for regulating and controlling rice resistance and application of the Ustilaginoidea virens gene UvASE2, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the Ustilaginoidea virens gene UvASE2 is as shown in SEQ ID NO: 1, and the amino acid sequence of the encoding protein of the Ustilaginoidea virens gene UvASE2 is as shown in SEQ ID NO: 2. By constructing a transgenic rice plant for heterologous expression of UvASE2, it is found that the transgenic rice can significantly improve the resistance to false smut and bacterial blight and significantly induce the outbreak of active oxygen. The heterologous expression UvASE2 gene of Ustilaginoidea virens has the function of positively regulating and controlling the disease resistance of rice, and the UvASE2 gene can be used for improving the disease resistance of rice and has important significance on creation of disease-resistant germplasm of rice.

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 present invention provides a novel process for the production of aliphatic unsaturated C10-aldehyde compounds catalyzed by lipoxygenase (LOX) from a polyunsaturated fatty acid (PUFA) source. The invention also relates to the isolation and characterization of novel, preferably bifunctional LOXs from different algae sources, and the identification of structural and / or functional related LOXs from different bacterial sources. The invention also relates to the provision of enzyme mutants derived from said newly identified enzymes. Another aspect of the invention relates to the corresponding coding sequences of said enzymes, recombinant vectors and recombinant host cells, which are suitable for the production of such LOXs and for carrying out a new process for the production of aliphatic unsaturated C10-aldehyde compounds. Another aspect of the invention relates to the use of a specific aldehyde or aldehyde mixture obtained according to the invention as a flavoring ingredient or an ingredient of a food or feed composition.

The invention discloses acyltransferase for synthesizing a polyester intermediate and application of the acyltransferase. The acyltransferase is specifically a wild-type enzyme with an amino acid sequence as shown in SEQ ID NO: 1 or an acyltransferase variant which is obtained by mutation of the wild-type enzyme and has acyl transfer reaction catalysis activity, and the acyltransferase variant is specifically obtained by replacing at least one amino acid residue in the 209th site, the 166th site and the 119th site in the amino acid sequence of the wild-type enzyme. And the protein with improved acyltransferase activity is obtained. The wild type enzyme and the acyltransferase variant provided by the invention both have acyltransferase activity. The method has an important application value.