131results about How to "Improve expression level" patented technology

The invention provides an application of a PavERF110 gene in regulating and controlling the ripening direction of sweet cherry fruits, and belongs to the technical field of molecular breeding. The nucleotide sequence of the PavERF110 gene is as shown in SEQ ID NO.1, and the amino acid sequence of the encoded protein of the PavERF110 gene is as shown in SEQ ID NO.2. The invention also provides an overexpression vector and a VIGS gene silencing vector constructed by the gene, a construction method of the overexpression vector and the VIGS gene silencing vector, and a recombinant microorganism containing the vectors. Engineering bacteria containing related carriers are injected and infected at the equator of the fruits 25 days after the sweet cherries blossom, and it is found that overexpression of the PavERF110 gene can accelerate fruit coloring and increase the expression level of the mature related genes. New technical support and theoretical basis are provided for sweet cherry fruit quality improvement and ripening regulation, market requirements can be met, and meanwhile understanding of a fruit ripening transcription regulation mechanism is deepened.

The invention provides a respiratory syncytial virus antigenic polypeptide, nucleic acid and a vaccine. Compared with the respiratory syncytial virus wild type Pre-F protein, the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment of the respiratory syncytial virus antigenic polypeptide has the following changes: the 104th to 144th regions of the Pre-F protein are replaced with GS linker; two groups of disulfide bonds are introduced into S155C, S290C, A149C and Y458C; the S190F and the V207L are used for realizing cavity filling; and P102A, L373R, I379V, M447V, P102A, L373R, the C terminal of the respiratory syncytial virus antigenic polypeptide or the immunogenic fragment thereof comprises a cytoplasm tail region fragment, and the cytoplasm tail region fragment is a fragment derived from 3-10 continuous amino acid residues of a PreF protein cytoplasm tail region. According to the present invention, the pre-F immunogenicity and / or stability can be improved, and the protein expression level can be easily improved.

The invention discloses a poplar PdbHLH57 gene as well as an expression protein and application thereof, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the poplar PdbHLH57 gene disclosed by the invention is as shown in SEQ ID NO.1, and the amino acid sequence of the expression protein of the poplar PdbHLH57 gene is as shown in SEQ ID NO.2. The method comprises the following steps: constructing an expression vector of a poplar PdbHLH57 gene, and transforming the expression vector into a poplar leaf; and cultivating and screening to obtain the transgenic poplar with red leaves. Results show that compared with wild poplar, the PdbHLH57 gene for promoting poplar anthocyanin synthesis has the advantages that transgenic poplar leaves become red, and the anthocyanin content in transgenic plant leaves is remarkably increased; the expression level of anthocyanin biosynthesis related genes is remarkably increased, and further, it is shown that the PdbHLH57 gene plays an important role in poplar anthocyanin biosynthesis.

The invention relates to application of cembratrienol in preparation of a medicine for preventing and / or treating psoriasis, and belongs to the technical field of biological medicine. The invention provides an application of cembratrienol in preparation of a medicine for preventing / or treating psoriasis. Researches find that cembratrienol can significantly inhibit secretion of inflammatory factors in a psoriasis-like cell model induced by M5; moreover, research finds that cembratrienol can significantly improve the skin lesion condition of an imiquimod-induced psoriasis mouse model, improve the weight loss degree of the psoriasis mouse model, relieve the pruritus condition of the psoriasis mouse model, improve the skin inflammation of the psoriasis mouse model, repair the skin barrier of the psoriasis mouse model, and improve the curative effect of the psoriasis mouse model. The body immune response of the psoriasis mouse model is reduced. Therefore, cembratrienol has a great application prospect in preparation of the medicine for preventing / treating psoriasis.

The invention discloses an rhFN recombinant human chemical fibronectin process of a genetic engineering technology, and particularly relates to the technical field of genetic engineering, and the rhFN recombinant human chemical fibronectin process comprises target gene acquisition, gene cloning and vector construction, host cell selection and transformation, cell culture and screening, protein expression and optimization, and protein separation and purification. According to the rhFN recombinant human chemical fibronectin process of the genetic engineering technology provided by the invention, high-quality target genes are accurately obtained through technologies such as new-generation sequencing and gene synthesis, and the gene expression efficiency and stability are enhanced by virtue of optimized vector construction and regulatory elements; protein expression ability and specificity are improved in host cell selection and transformation, efficient culture and accurate screening are realized in cell culture and screening links, total yield is increased in a protein expression optimization link, purity and efficiency are improved in a protein separation and purification link, and product quality and safety are ensured through comprehensive quality control. The requirements in the fields of medical treatment and scientific research are expected to be better met.

The invention discloses a mutant with improved enzyme activity of sucrose isomerase and application of the mutant, and belongs to the technical field of genetic engineering and enzyme engineering in molecular biology. A known sucrose isomerase parent is subjected to molecular modification by utilizing a site-directed mutagenesis technology to obtain mutant enzymes T522S / K340D, D222H / K340D, G467N / K340D, T522S, K340D, D222H and G467N, and the catalytic activity of the series of mutants provided by the invention is increased by 10%-25% compared with that of original bacteria; the isomaltulose conversion rates of the combined mutants T522S / K340D, D222H / K340D and G467N / K340D are respectively increased by 2.3%, 1.5% and 1.3% compared with those of original bacteria, and the strain has good stability, can be used for biological preparation of isomaltulose and has potential application prospects.

The invention discloses lnc.3024.2-GbMYB2 for regulating and controlling synthesis of gingko flavonoids and application of the lnc.3024.2-GbMYB2, and belongs to the technical field of genetic engineering, aiming at analyzing the biological functions of lncRNA (lnc.3024.2) and MYB target gene (Gb06719) participating in gingko flavonoids anabolism, by utilizing the technical means of bioinformatics analysis, real-time fluorescence quantification, subcellular localization, prokaryotic expression, genetic transformation and the like, the GbMYB2 can be used for regulating and controlling synthesis of the gingko flavonoids. The regulation mechanism of the transcription factor lnc.3024.2 and the target gene GbMYB2 of the lnc.3024.2 participating in the synthesis of the flavonoids in the ginkgo leaves is discussed. And a gene regulation network model of gingko flavonoid anabolism is further enriched. On the basis, the molecular mechanism of regulating and controlling the synthesis of the gingko flavonoids by the lncRNA targeting MYB is further analyzed on the early-stage basis, the content of the gingko flavonoids is increased by utilizing a genetic engineering technical means, and a new way is provided for researching the synthesis way of the gingko flavonoids.

The invention relates to the technical field of biology, in particular to recombinant bacillus for heterologous synthesis of ovotransferrin as well as a construction method and application of the recombinant bacillus. According to the invention, bacillus (bacillus subtilis and bacillus amyloliquefaciens) is taken as a host, and the OVT gene is heterologously expressed by using a genetic engineering technology, so that heterologous secretory expression of OVT1 and OVT2 from different sources in chassis cells is realized; meanwhile, the invention also provides a method for more economically and efficiently increasing the expression level of ovotransferrin. Three different molecular chaperones PrsA, DnaK and CasA are further selected for co-expression with the ovotransferrin, correct folding of the ovotransferrin in host cells is promoted, the soluble secretory expression quantity of the ovotransferrin is increased, and the expression efficiency of the ovotransferrin is improved.

The invention discloses a construction method of a heparin 2-O-sulfotransferase high-activity mutant, and belongs to the technical field of biology. According to the heparin 2-O-sulfotransferase and the application of the heparin 2-O-sulfotransferase, a dissolution promoting label is fused at the N end of the heparin 2-O-sulfotransferase, the coding sequence of the dissolution promoting label is optimized, and active expression of the heparin 2-O-sulfotransferase is achieved; according to the invention, the heparin 2-O-sulfotransferase is mutated, so that a mutant with high stability and catalytic activity is obtained; furthermore, fermentation conditions of the heparin 2-O-sulfotransferase are optimized, so that the expression quantity and the enzyme activity are improved, and the application potential of the heparin 2-O-sulfotransferase in industrial production is enhanced.

The invention provides application of baicalin in preparation of a medicine for treating streptococcus agalactiae induced mastitis, and relates to the technical field of medicine. A mouse is selected as an animal model to research the intervention effect and mechanism of the baicalin on the streptococcus agalactiae induced mouse mastitis inflammatory response, and it is clear that the baicalin can play a role in protecting the mouse mammary gland by regulating and controlling an NLRP3 inflammatory small body pathway; a reference basis is provided for treating the cow mastitis caused by streptococcus agalactiae infection by applying the baicalin.

The invention provides an application of a PavMYB1 gene in regulation and control of sweet cherry anthocyanin synthesis. The nucleotide sequence of the PavMYB1 gene is as shown in SEQ ID NO. 1, and the amino acid sequence of the encoded protein of the PavMYB1 gene is as shown in SEQ ID NO. 2. The invention provides an overexpression vector and a VIGS gene silencing vector constructed by related genes, a construction method of the overexpression vector and the VIGS gene silencing vector, and a recombinant microorganism containing the vectors. Experiments prove that overexpression of PavMYB1 can increase fruit pigment accumulation and anthocyanin synthesis key gene expression, technical support and theoretical basis are provided for improvement of sweet cherry fruit quality, and revealing of a regulation mechanism of sweet cherry anthocyanin synthesis is facilitated.

The invention relates to the technical field of plant genetic engineering, and provides an agrobacterium rhizogenes-mediated sunflower transient transformation system and a construction method thereof, sterile conditions and tissue culture are not needed, a vector to be transformed is transferred into agrobacterium rhizogenes, positive strains are cultured and screened, and a dip dyeing solution is prepared; cutting a stem section of the sunflower, and pricking holes near a cut; immersing the stem segments in the dip dyeing liquid for a period of time; taking out and inserting into sterilized vermiculite soil mixed with orobanche coerulescens for rooting to obtain an instantly transformed sunflower plant; the method is easy and convenient to operate and short in period, the expression level of the HaCERK1 gene can be effectively improved, the resistance of sunflowers to orobanche coerulescens infection is enhanced, and the gene function research period is shortened.

The invention provides a high-yield L-cysteine engineering bacterium accumulated by sulfur donor 3 '-phosphoadenosine-5'-phosphoryl sulfuric acid as well as a construction method and application of the high-yield L-cysteine engineering bacterium. According to the method, the copy number of key genes in a cysteine synthesis route in an escherichia coli organism is changed, heterologous genes are introduced, the expression level of the key genes in the cysteine synthesis route in the escherichia coli organism is enhanced, and sulfur flux is further made to flow to synthesis of L-cysteine. On the basis that the chassis bacteria enhance the carbon flux, in order to maintain the balance of carbon and sulfur in the strain and continuously enhance the synthesis of the sulfur flux flowing to the L-cysteine, the synthesis of the product is promoted by enhancing the synthesis capability of a sulfur donor PAPS in a sulfate pathway, and finally, the plasmid-free and antibiotic-free engineering strain for high yield of the L-cysteine is obtained. Finally, the shake flask titer of the L-cysteine reaches 6.06 g / L and is increased by 49.2% compared with that of an original strain.

The invention belongs to the technical field of biological medicines, and discloses application of a ULK1 agonist in preparation of an anti-depression medicine and application of ULK1 in preparation of a depression diagnosis product. Experiments show that ULK1 not only can be used as a biomarker for diagnosing depression, but also can be used as an important target for developing drugs for treating / preventing depression. In addition, the invention further indicates that the ULK1 agonist BL-918 has a rapid anti-depression effect and can be used for treating / preventing depression.

The invention provides a plant cytosine single-base editor as well as a construction method and application thereof, and relates to the technical field of plant genetic engineering. The single-base editor provided by the invention is named as hyPopCBE, and has a structural formula shown in the description, the editor comprises hA3A / Y130F cytosine deaminase, an optimized human-derived Rab51 single-stranded DNA binding protein structural domain, a fusion protein of nCas9 nuclease and a uracil glycosylase inhibitor, a fusion protein expression unit of phage MS2 coat protein and the uracil glycosylase inhibitor, and a guide RNA transcription expression unit integrated with an MS2 binding sequence. Compared with an existing plant cytosine single-base editor, the single-base editor constructed by the method can remarkably improve the editing efficiency of plant alleles, and the stability is higher; when the herbicide-resistant poplar gene is applied to poplar, a new herbicide-resistant poplar germplasm can be successfully created, and a technical support is provided for research and molecular breeding of a poplar functional genome.

The invention relates to the field of genetic engineering, in particular to an optimized bacillus subtilis expression vector, a genetically engineered bacterium containing the expression vector and application of the genetically engineered bacterium. The vector is based on pMA5 plasmids, and a shuttle type bacillus subtilis expression vector is optimized; an open reading framework with unknown functions in the bacillus subtilis expression vector is reduced, so that the size of the vector is reduced, the expression burden of cells is reduced, and an optimization effect is achieved; the optimized bacillus subtilis expression vector not only can be stably copied in escherichia coli, but also can be stably expressed in bacillus subtilis, and the expression level of heterologous gene coding protein is improved by utilizing the genetically engineered bacteria containing the optimized bacillus subtilis expression vector.

The invention relates to the technical field of plant genetics, in particular to a TTG1 protein and a coding gene and application thereof. The TTG1 protein comprises an amino acid sequence as shown in SEQ ID NO. 1. Based on full-length transcriptome data of pennisetum alopecuroides, the TTG1 protein and the encoding gene thereof are obtained through research. The TTG1 protein provided by the invention not only can effectively regulate and control the anthocyanin content in plants, but also can regulate and control the development of leaves, furs and hairs of the plants. The improved TTG1 protein can be used for cultivating high-resistance pennisetum alopecuroides varieties, and has important significance in the field of developing high-quality pennisetum alopecuroides varieties.

The invention discloses application of Axin1 in regulation and control of osteogenic differentiation of dental pulp stem cells of deciduous teeth. The research shows that the overexpression and knock-down of Axin1 do not influence the proliferation of SHED and the characteristics of MSC. However, overexpression of Axin1 promotes migration of SHED, and this effect can be weakened by Axin1 silencing. Overexpression of Axin1 promotes osteogenic differentiation of SHED, and knock-down of Axin1 shows a reverse effect. In mechanism, overexpression of Axin1 obviously improves the expression level of phosphorylated ERK1 / 2, so that osteogenic differentiation of SHED is promoted. The promotion effect of overexpression of Axin1 on osteogenic differentiation of SHED can be eliminated by using an ERK1 / 2 chemical inhibitor for treatment, which indicates that Axin1 plays a role as a key regulatory factor for osteogenic differentiation in SHED through an ERK1 / 2 pathway. A research result shows that Axin1 can be used as a potential new target and is used for treating diseases related to bone regeneration by applying SHED.

The application belongs to the technical field of biological medicine, and particularly relates to a kind of lipid nanoparticles and engineered immune cells. The application first provides an engineered immune cell, the engineered immune cell includes immune cell, lipid nanoparticle and exogenous drug;The exogenous drug is loaded in the lipid nanoparticle;When the immune cell is contacted with the lipid nanoparticle, the immune cell and the lipid nanoparticle endocytosis occur to obtain the engineered immune cell. In another aspect, the application also provides a kind of lipid nanoparticles that can improve the encapsulation efficiency of mRNA and the delivery efficiency of immune cells.

The invention provides application of adrenochromic hydrazone in preparation of a medicine for treating subdural hematoma, and belongs to the technical field of medicines. The invention finds that the adrenochrome can improve the expression level of endothelial connexin of epidural blood vessels, enhance the resistance of blood capillaries to injury, reduce the permeability of the blood capillaries, reduce the bleeding amount of subdural hematoma, relieve inflammatory response caused by hematoma stimulation and accelerate the recovery of subdural hematoma for the first time. Therefore, the compound can be used for preparing medicines for treating subdural hematoma. The medicine can be administrated through oral administration, the risk existing in a traditional drilling drainage operation is avoided, and an alternative or auxiliary treatment scheme is provided for patients who are intolerant to statins or hormones and patients who are ineffective in treatment through combination of statins and hormones.

The invention relates to a traditional Chinese medicine composition for treating chronic kidney diseases and application thereof. The traditional Chinese medicine composition is prepared from the following traditional Chinese medicine raw materials: astragalus membranaceus, rheum officinale, cassia twig, poria cocos, radix paeoniae rubra, peach kernels and moutan bark. The traditional Chinese medicine components in the traditional Chinese medicine composition are scientific in compatibility, and the traditional Chinese medicine composition has a very remarkable effect on treating the chronic kidney diseases. Researches show that the traditional Chinese medicine composition can effectively treat the chronic kidney disease, improve the kidney function of a kidney disease patient, reduce the serum creatinine level and the urea nitrogen level, increase the expression level of peroxisome related functional enzymes and relieve kidney pathological injuries.

The present invention provides a tight junction protein antibody H04 and its application, wherein the antibody H04 is a single-domain antibody, and the heavy chain variable region of the single-domain antibody includes CDR1, CDR2 and CDR3; the amino acid sequence of the CDR1 includes SEQ ID NO.1; the amino acid sequence of the CDR2 includes SEQ ID NO.2; and the amino acid sequence of the CDR3 includes SEQ ID NO.3. The tight junction protein antibody H04 obtained by the present invention has a higher affinity for CLDN6 and a lower affinity for CLDN9, that is, it has a higher specificity for CLDN6, and at the same time, the tumor killing effect of ADCC and CAR-T mediated by it is better, and the chimeric antigen receptor molecule constructed based on it can better mediate immune cell killing of tumors.

The invention relates to the field of biological medicine, and provides an ionizable lipid compound capable of activating an STING signal channel and application of the ionizable lipid compound. The invention specifically provides an ionizable lipid compound which has a natural adjuvant effect and can induce generation of I-type interferon gene expression by activating an STING pathway, so that APC activation and antigen presentation are promoted, and the immune response ability of a body is enhanced. And a method for synthesizing the ionizable lipid compound from a ketone compound, an amine compound and an isonitrile compound. The invention also provides an application of the ionizable lipid compound in preparation of lipid nanoparticles, the lipid nanoparticles formed by the ionizable lipid can deliver nucleic acid molecules into cells, the expression level of nucleic acid drugs in vivo can be significantly improved, and the immune response of the organism can be stimulated. Ionizable lipid compound having the structure shown: # imgabs0 #

The present invention belongs to the field of microorganisms and biomedicine technology, and specifically relates to an olive-colored chaetomium histone deacetylase and its encoding gene and application. The present invention performs gene knockout on HDAC in the fungal strain olive-colored chaetomium (Chaetomium olivaceum) SD-80A, realizes the activation of the recessive biosynthetic gene cluster and increases the yield of the compound cochlidinol. Studies have found that the compound shows good antimicrobial, anti-inflammatory, anticoagulant and anticancer activities, has the potential to be developed as a drug lead compound, and therefore has good practical application value.

The present application relates to the field of cell therapy and regenerative medicine, and in particular to a kind of old age adipose tissue-derived mesenchymal stem cells pretreated by MK8722, its preparation method and application.The method comprises the following steps: the old age adipose tissue-derived mesenchymal stem cells are inoculated in culture solution containing MK8722 to carry out in vitro treatment, and first cells are obtained;The culture solution containing MK8722 is removed, and the first cells are washed to obtain second cells;Second cells are cultured using culture solution without MK8722, and the old age adipose tissue-derived mesenchymal stem cells pretreated by MK8722 are obtained.The cells obtained by the method, relative to the old age adipose tissue-derived mesenchymal stem cells without the in vitro treatment, show at least one improvement in proliferation capacity, continuous passage amplification capacity, adipogenic differentiation capacity or chondrogenic differentiation capacity, and can be used to prepare products for treating or alleviating osteoarthritis.

The invention relates to an RNA (Ribonucleic Acid) coding element and a method for carrying out gene editing on a plant by using the RNA coding element. In particular to an RNA (Ribonucleic Acid) coding element, an sgRNA bidirectional enhanced expression element, a carrier capable of carrying out efficient gene editing in a plant and a method for carrying out gene editing on the plant. The vector can improve the gene editing efficiency and the targeting specificity of the plant, and the sgRNA bidirectional enhanced expression sequence can further improve the gene editing efficiency and the targeting specificity of the plant.

The invention relates to the technical field of preparation of microbial enzyme preparations, in particular to a microbial enzyme preparation for polyester plastic degradation and a preparation method of the microbial enzyme preparation. The present invention relates to a microbial composition comprising at least one microbial strain engineered to enhance PETase expression and maintain activity under hypoxic conditions, and a nanocarrier, and also comprising chitosan. The expression level of microorganisms on PETase is enhanced through gene modification, the degradation efficiency of polyester plastics is improved, a nano-carrier material and a chitosan protective film are introduced, the stability and bioavailability of the enzyme preparation are improved, and the prepared microbial enzyme preparation can keep activity in a low-oxygen, illumination or complex environment, and can be used for preparing a biological enzyme preparation. And secondly, the whole preparation process does not depend on harmful chemicals, the green chemistry principle is met, and environmental protection and resource recycling are facilitated.

The invention relates to a 3 'UTR (Untranslated Region) sequence for promoting RNA (Ribonucleic Acid) expression and application of the 3' UTR sequence in mRNA (Messenger Ribonucleic Acid) medicines. The 3 'UTR sequence comprises polynucleotide containing any one of SEQ ID NO.1-37. The 3 'UTR provided by the invention has a stronger function of promoting mRNA expression. The 3 'UTR fragment provided by the invention plays a role in promoting translation in different types of host cells, has broad spectrum in action, and shows an application prospect in RNA (Ribonucleic Acid) therapeutic drugs or mRNA (Messenger Ribonucleic Acid) vaccines. The 3 'UTR obtained through optimization has a better target protein expression effect and can be used for mRNA vaccine development or RNA therapeutic drugs.

The application discloses a method for culturing three-dimensional cell spheres of alveolar type II epithelial cells under a condition without exogenous matrix glue, and comprises the following steps: A, perfusion digestion treatment is performed on mouse lung tissue to obtain a single cell suspension; B, the single cell suspension is inoculated into a culture dish coated with anti-CD31 antibody and anti-CD45 antibody for immunoadhesion culture, under the culture condition, cells expressing CD31 and CD45 are subjected to immunoadhesion, meanwhile, fibroblasts with strong adhesion ability are attached to the surface of the culture dish, so that non-epithelial cells are removed, and unattached cells are collected to obtain an enriched ATII cell population; and C, the unattached cells are inoculated into a low-adhesion culture environment for suspension culture, and the culture system contains signal regulatory factors for maintaining the stability of the ATII cell phenotype, and the enrichment degree of epithelial cells is improved, and the interference of non-target cells is reduced through the combination of negative immunoadhesion of the anti-CD31 antibody and the anti-CD45 antibody and short-time adhesion treatment.

The invention belongs to the technical field of medicines, and particularly provides a novel application of methyl cedryl ether in preparation of a product for resisting skin photoaging. Cell experiment evaluation proves that the methyl cedryl ether has no obvious influence on the activity of dermal Hs68 fibroblasts at the concentration of 12.5-100 mu M, and meanwhile, the methyl cedryl ether can obviously increase the content of hyaluronic acid in light-aged Hs68 cells induced by medium-wave ultraviolet rays (UVB), improve the mRNA expression level of COL1A2 and COL3A1, improve the mRNA expression level of HAS1 and HAS3 and reduce the mRNA expression level of MMP-1, MMP-3 and MMP-9. Therefore, the methyl cedryl ether provided by the invention can be applied to preparation of products for resisting skin photoaging.