64 results about "Vacuole" patented technology

The invention relates to recombinant pichia pastoris for high expression of porcine pancreatic phospholipase A2 and application of the recombinant pichia pastoris, and belongs to the technical field of genetic engineering. According to the method disclosed by the invention, a vacuole sorting receptor Vps10 is knocked out by utilizing a CRISPR-Cas9 system, a CPY sorting way between a trans-golgi apparatus and a vacuole is interrupted, and ppPLA2 degradation is reduced; then, in order to relieve unfolded protein response in the endoplasmic reticulum, molecular chaperones are introduced into the genome, and synthesis of ppPLA2 is enhanced; the expression intensity of the ppPLA2 is improved by increasing the gene copy number of the ppPLA2; and finally, high-density fermentation is performed in a 3-L bioreactor, so that high-efficiency expression of the ppPLA2 in the pichia pastoris is realized.

The invention provides an application of an OsINV2 gene in regulation and control of rice cold stress resistance. A nucleotide sequence of the OsINV2 is shown as SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. By revealing the key regulation and control effect of the vacuole sucrose invertase gene OsINV2 in rice low-temperature stress response, the molecular mechanism that the gene affects the cold resistance of plants by regulating the sugar metabolism balance in vacuoles is illustrated for the first time. The expression of the OsINV2 is regulated in a targeted manner through molecular marker-assisted breeding or a gene editing technology, so that the low-temperature survival rate in the seedling stage and the cold resistance in the booting stage of the rice can be remarkably improved, glumous flower degeneration and pollen abortion caused by low temperature are reduced, and meanwhile, the limitation of the cold resistance of varieties on geographical distribution is broken through; a theoretical support and a technical path are provided for breeding and adaptive popularization of cold-resistant rice varieties.

The invention discloses a single vesicle particle electrochemical signal identification and classification method and identification system, and the method comprises the steps: S1, preparing a carbon fiber ultramicro electrode, applying a voltage in a patch clamp system constant potential mode, and collecting picoampere-level current response generated by oxidation of a single vesicle content on the surface of the electrode; s2, carrying out smooth filtering on the collected original current signal, and realizing characteristic peak identification and multi-dimensional physical / statistical characteristic extraction by using a multi-parameter peak identification method based on a threshold value; s3, carrying out preprocessing operations including missing value processing, standardization, principal component analysis dimension reduction and category equalization on the extracted peak value characteristic data; and S4, on the basis of the preprocessed feature data, constructing and training at least one machine learning model for automatically identifying the particle size category of the vesicle particles. The method combines the advantages of single-vesicle electrochemical high-sensitivity measurement and intelligent algorithm classification, and has the advantages of high recognition efficiency, good repeatability, wide applicability and the like.

A genetically modified yeast cell, wherein the yeast cell comprises a genetic modification comprising overexpression of yeast gene encoding porphobilinogen deaminase (HEM3), the HEM3 gene having at least 80% identity with SEQ ID No. 7. The genome of the modified yeast cell further comprises one or more genetic modifications in one or more genes selected from: genes coding for heme-dependent repressor of hypoxic genes (ROX1), genes coding for heme oxygenase (HMX1), genes coding for a receptor for vacuolar proteases (VPS10), and genes coding for vacuolar proteinase (PEP4), the one or more genetic modifications being such that expression of a polypeptide from such a gene is reduced or disrupted or the polypeptide expressed is non-functional.

The invention discloses a method for preventing and controlling cadmium accumulation of rice grains and strengthening zinc concentration at the same time, and relates to the technical field of agricultural environment and crop nutrition regulation, the single regulation defect is precisely overcome through the synergistic effect of a compound foliage regulator, salicylic acid and a zinc amino acid chelate are compounded to form the regulator, and the regulation effect is improved. Salicylic acid is utilized to induce synthesis of plant chelating peptide in rice, cadmium vacuole partitioning is enhanced, cadmium transporter protein expression is reduced, and migration of cadmium to grains is prevented and controlled from a transport path; meanwhile, zinc is rapidly supplemented by means of the efficient absorption characteristic of the zinc amino acid chelate, and the zinc amino acid chelate can cooperate with a salicylic acid signal to further inhibit cadmium distribution, so that the problems that a single zinc fertilizer cannot resist cadmium and single salicylic acid cannot be rich in zinc are solved, cadmium resistance and zinc enrichment are synchronously promoted, and meanwhile, dry-wet alternate irrigation in the booting stage and the mature stage is matched, so that the yield is increased. By regulating the soil moisture state, the physiological metabolism environment of the rice is optimized, the absorption and utilization of a regulating agent are promoted, and the vacuole partitioning effect of cadmium and the phloem transfer efficiency of zinc are enhanced.

The invention discloses a synbiotics-like feed prepared from vegetable waste polymerized vegetable cakes and a method, and relates to the field of agricultural circular economy, cellulose in vegetable waste cell walls is effectively decomposed under mild conditions through a specific catalytic decomposition enzyme combination, and part of cellulose is converted into carbon dioxide and water, so that the utilization rate of the cellulose is improved. By combining physical heat energy conduction, liquid in cell vesicles is heated to generate micro air pressure to break through residual cell walls, so that recessive water in cells is successfully converted into surface dominant water, the energy consumption in the dehydration process is greatly reduced, and the loss of dry basis substances in the resource utilization process is effectively reduced; the vegetable cake product obtained through final polymerization is rich in various nutrient substances such as crude fibers and / or dietary fibers, proteins, vitamin C and carbohydrates, the recycling rate of the vegetable waste is larger than or equal to 95%, the recycling efficiency and the additional value are remarkably improved, meanwhile, the difficulty of subsequent harmless treatment is reduced from the source, and the economic input and the treatment cost are reduced.

The invention discloses application of TPN171H, an active fragment thereof or a derivative thereof in preparation of a medicine for treating spinal cord injury. The TPN171H shows an excellent treatment effect in spinal cord injury treatment, the levels of apoptosis factors BAX and Caspase-3 cleaved can be remarkably reduced, the level of Bcl-2 is improved, the content of a neuron marker MAP-2 is also remarkably increased, and the number of Tunel bodies and the cavitation-like structure are remarkably reduced. The intervention of the TPN171H also can reduce MNSS, and prompts that the TPN171H has the functions of promoting the spinal cord movement and coordination of the rat with spinal cord injury and improving the sensory system of the rat with spinal cord injury.

The invention belongs to the technical field of genetic engineering and biology, and relates to application of trichoderma harzianum TrVPT1 in regulation and control of trichoderma phosphorus accumulation and gibberellic disease resistance. The nucleotide sequence of the trichoderma harzianum vacuole phosphate transporter gene TrVPT1 is as shown in SEQ ID NO: 1. The invention proves that the TrVPT1 gene has an important regulation and control effect in trichoderma phosphorus steady-state regulation and control and biological prevention and control functions.

This invention relates to the field of anti-aging drug technology, specifically to the application of small molecule compounds in the preparation of anti-aging drugs. These small molecule compounds competitively block the interaction between KDM6A and the vacuolar ATPase V0 domain subunit ATP6V0C by targeting a specific small molecule binding pocket of the histone demethylase KDM6A, thereby inhibiting the demethylation activity of KDM6A on lysine residue 36 of ATP6V0C. These small molecule compounds can effectively downregulate aging-related β-galactosidase activity, improve cell cycle arrest, and reverse the senescent phenotype of nerve cells induced by the environmental pollutant GenX, making them particularly suitable for combating premature aging of nerve cells induced by perfluorinated and polyfluoroalkyl compounds such as GenX. This technical solution can solve the technical problem of the lack of drugs with clearly defined components, a clear mechanism of action, stable efficacy, and the ability to specifically alleviate aging in existing technologies, and has ideal prospects for widespread application.

The invention relates to the technical field of anti-aging drugs, in particular to application of a small molecule compound in preparation of anti-aging drugs. The small molecule compound competitively blocks the interaction between the KDM6A and a vacuolar ATP enzyme V0 structural domain subunit ATP6V0C through a specific small molecule binding pocket of targeted histone demethylase KDM6A, and further inhibits the demethylation activity of the KDM6A on 36th lysine of the ATP6V0C. The small molecule compound can effectively down-regulate the activity of senescence-related beta-galactosidase, improve cell cycle arrest and reverse nerve cell senescence phenotypes induced by environmental pollutant GenX, and is especially suitable for resisting nerve cell premature senescence caused by perfluorinated and polyfluoroalkyl compounds such as GenX. According to the technical scheme, the technical problem that drugs which are definite in component, clear in action mechanism, stable in curative effect and capable of pointedly relieving senescence are lacked in the prior art can be solved, and ideal application and popularization prospects are achieved.

The invention discloses a preparation method of a coronary artery microcirculation disturbance animal model, and belongs to the technical field of biological medicine. According to the method, the stable mouse CMD model can be established by injecting 20-40 [mu] g / mL Calcimycin into the cardiac apex for the first time. Calcimycin is a calcium ion (Ca < 2 + >) carrier commonly used in a laboratory and can transport calcium ions outside cells into the cells, so that the concentration of the calcium ions in the cells is increased. Particularly, when 40 [mu] g / mL of Calcimycin is injected, the pathological severity of the Calcimycin is further aggravated, myocardial fiber arrangement is disordered, boundary is unclear, a large number of myocardial cells are necrotic and dissolved, cavitation is formed, vascular wall degeneration swelling is caused, inflammatory cell infiltration is caused, and thrombus is formed in microvessels.

This invention discloses a genetically engineered bacterium for synthesizing carotenoids, its construction method, and its applications, belonging to the field of genetic engineering technology. The genetically engineered bacterium uses a carotenoid-producing yeast strain as the starting material. The gene ROX1, encoding a heme-dependent hypoxia gene repressor, is knocked out, while the expression of genes STB5 (encoding a transcription factor involved in NADPH regeneration), DID2 (encoding a class E protein in the vacuole protein sorting pathway), and VOA1 (encoding an endoplasmic reticulum protein playing a role in the assembly of the V0 region of V-ATPase) is upregulated. This invention promotes carotenoid synthesis and significantly increases carotenoid yield through combined regulation of gene expression outside the carotenoid synthesis pathway, demonstrating promising application prospects.

The invention discloses an application of luteolin-7-diglucuronide in preparation of a medicine for resisting liver fibrosis related diseases. According to the invention, two animal models, namely a CCl4-induced hepatic fibrosis mouse model and a high-fat, high-cholesterol and high-fructose (GAN) diet and CCl4-induced NASH mouse, are adopted; the results respectively prove that the luteolin-7-diglucuronide has obvious inhibition of mouse collagen deposition and exerts anti-hepatic fibrosis activity on CCl4 induced hepatic fibrosis mice and GAN diet combined CCl4 induced NASH mouse models. Besides, the luteolin-7-diglucuronide is proved to be capable of remarkably inhibiting pathological phenotypes such as formation of fat vacuoles, inflammatory cell infiltration and the like of NASH mice through a GAN diet and CCl4 induced NASH mouse model, and the luteolin-7-diglucuronide is proved to have an application prospect of being developed into an anti-NASH medicine.

The invention relates to the technical field of microbial fermentation and genetic engineering, and particularly discloses a yeast engineering bacterium for efficiently producing plastic depolymerases and application of the yeast engineering bacterium. The engineering yeast takes pichia pastoris as an expression host, a molecular chaperone is co-expressed, a vacuole sorting receptor gene is knocked out or knocked down, in addition, a yeast cell factory for efficiently expressing the plastic depolymerases is constructed by further combining regulation and control element optimization and target gene copy number regulation, and the engineering yeast has the advantages that the engineering yeast can be used for efficiently expressing the plastic depolymerases; the regulatory element comprises any one or a combination of more of a strong promoter, a signal peptide and a C-terminal tag. The engineering yeast provided by the invention can realize efficient accumulation and secretion of the plastic depolymerases, obviously improve the enzyme yield and reduce the production cost of unit enzyme activity, and provides a stable and efficient yeast expression system for industrial and low-cost production of the plastic depolymerases; the method has an important application prospect in the fields of biodegradation and resource utilization of waste plastics.

The present application relates to lipid vacuole (aka LV)-containing lipocartilage cells (aka LCs), and methods for the differentiation of human lipocartilage progenitors into LV-containing LCs. Also disclosed are methods related to purifying, labeling, maturing, and delivering of LV-containing LCs to a subject in need thereof.

Methods and compositions are provided for enhancing the accumulation and / or recovery of recombinant casein expressed in soy. Endogenous Casein-Processing Interfering Proteins (CPIPs), such as oxidases like lipoxygenases (LOX), are present in the recombinant protein fractions and interfere with yield and / or purity. RNA interference (RNAi) mechanisms are leveraged in plasmids that knockdown genes expressing CPIPs and / or genes in the Soybean Vesicular Trafficking Pathway (SVTP). Knocking down of two or more target genes is manifested using a dual gene targeting approach, often via a single expression cassette containing multiple distinct shRNA constructs for enhanced efficiency. The shRNA constructs invoke a RNAi mechanism that leads to increased casein accumulation either: (1) directly, by suppressing CPIPs (e.g., LOX) to enable more efficient purification; or (2) indirectly, by modifying SVTP genes to route casein away from lytic vacuoles, preventing degradation.

The invention discloses staining agent uniform smearing equipment for a clinical laboratory, and belongs to the technical field of medical instruments, the staining agent uniform smearing equipment comprises an automatic smear conveying belt, the automatic smear conveying belt is provided with a plurality of smear placing mechanisms, each smear placing mechanism is provided with a bubble left-right swing removing mechanism, and the top of the automatic smear conveying belt is provided with a uniform smearing mechanism. And a bubble rolling removal mechanism is arranged on the automatic smear conveying belt. The bubble left-right swing removing mechanism works to push deep bubbles to the surface, the bubble rolling removing mechanism thoroughly discharges surface bubbles and corner-containing tiny bubbles, and for areas such as four corners and the edge of a smear where the bubble left-right swing removing mechanism cannot reach, due to the fact that stress on the edge of the smear is weak during swinging, tiny bubbles are prone to remaining, and the smear is not prone to falling off. The bubbles at the corners can be thoroughly removed through rolling coverage of the driving wheel, so that the surface of the smear is not blocked by any bubble, the situation of misjudgment of cell vacuoles cannot occur during subsequent microscope observation, and the blood cell morphology recognition accuracy is improved.

The invention discloses a biphase gel material formed based on hydrogen bond self-assembly and a preparation method and application thereof.The biphase gel material is formed by a first polymer and a second polymer based on hydrogen bond self-assembly, the first polymer comprises a branched polymer, the second polymer comprises a long-chain linear multi-hydrogen-bond site polymer, and the long-chain linear multi-hydrogen-bond site polymer comprises a long-chain linear multi-hydrogen-bond site polymer. The branched polymer comprises at least one of the following substances: 1) a polyether polymer with hydroxyl at the tail end; and 2) a polymer obtained by carrying out a derivatization reaction on the hydroxyl of the polyether polymer in the step 1). A biological inert film can be formed in situ on the surface of the water-loss gel, so that invasion of exogenous bacteria is effectively blocked. A skeleton-vacuole structure is formed inside due to microscopic phase separation, so that a large number of bacteria or cells and the like can be loaded. The unique internal and external structures of the two-phase gel material can load wide and complex contents and provide a steady state for the contents, and meanwhile, the two-phase gel material can be widely applied to skin diseases, implant materials and the like due to excellent mechanical properties of the two-phase gel material.

The invention discloses an application of two ciliate desert-grass AsIII reverse transport proteins in improving the arsenic hyperaccumulation capability of plants. The application is as follows. According to the invention, the PvACR3 of a cell membrane and the PvACR3 positioned in a vacuolar membrane are simultaneously subjected to heterologous overexpression in a plant through a hybridization technology; 1, the biomass of the plant is obviously improved, the hyperaccumulation capability of the plant on arsenic is enhanced, and the absorption and transport efficiency of the plant on arsenic is promoted; the invention provides an efficient, environment-friendly and low-cost new technical choice for the bioremediation technology of the arsenic-polluted soil.

The application provides a compound, a preparation method and application in preparation of a medicine for treating tumors, and relates to the technical field of chemical synthesis. The compound can stably induce cells to produce pyroptosis characteristic cell membrane large bubbles, simultaneously induce intracellular formation of a large vacuole-like structure similar to the characteristics of vacuole death and paraptosis, and has significant tumor cell proliferation inhibition activity, thereby providing a new chemical entity and research idea for research and development of a new type of anti-tumor medicine and research of new cell death mechanisms such as pyroptosis, vacuole death and paraptosis.

The present invention relates to a live attenuated Gram-negative bacterium comprising a modified hlyCABD operon, wherein the modified hlyCABD operon is split into a first segment and a second segment, the first segment being operably linked to a first independently controlled promoter, wherein the first independently controlled promoter is a strong constitutive promoter or a strong vacuole-induced promoter, and the second segment being operably linked to a second independently controlled promoter, wherein the second independently controlled promoter is a strong vacuole-induced promoter, and wherein the first segment comprises a heterologous polynucleotide encoding a lysin upstream of a hlyAs translocation sequence, wherein the heterologous polynucleotide encoding one or more cargo molecules replaces a hlyA gene, and wherein the second segment comprises hly genes involved in secretion.

The invention discloses a preparation method of rapeseed oil with high polyphenol retention rate based on bionic enzymolysis. Comprising the following steps: raw material pretreatment: crushing rapeseeds and adjusting the water content; enzymatic hydrolysis activation: spraying a beta-glucosidase solution, and standing to obtain pretreated rapeseeds; low-temperature squeezing: feeding the pretreated rapeseeds into a screw press, and mounting a bionic ceramic filter element at an oil outlet; and directly filling the rapeseed oil flowing out of the bionic ceramic filter element to obtain the rapeseed oil. The bionic ceramic filter element is sintered by adopting a gradient sintering technology, forms a pore channel structure which is dense outside and sparse inside, and mainly comprises three layers: an outer layer: 5-10 [mu] m macropores and simulates a cell wall screen structure; the middle layer is a pore channel of 1-3 microns and is filled with a compound enzyme-buffer gel solution with the pH value of 5.5; the inner layer is provided with 50-100nm micropores which are used for adsorbing phospholipid. According to the method, the microenvironment of plant cell vesicles is simulated, and triple bionic design of gradient porous ceramic + buffer gel + enzyme-substrate pre-combination is adopted, so that efficient and stable enzyme catalysis is realized, and the rapeseed oil with high polyphenol retention rate is obtained.

Compositions and methods for increasing fatty acid and / or oil accumulation in the stems of plants are provided. Constructs including stem cell-specific promoters operably linked to nucleic acids encoding a plastidic phosphoenolpyruvate / phosphate translocator protein and a tonoplast-localized hexose transporter protein are described.

The invention discloses a natural hirudin extraction method and application, and relates to the technical field of natural hirudin extraction.The method is characterized by comprising the steps that a tissue structure capable of being subjected to salivary gland single cell combined gland culture is selected; washing away bloodstain in the tissue structure by using PBS, then washing for three times by using the antibacterial liquid, and finally washing away the residual antibacterial liquid by using PBS; inoculating the tissue structure to a cell culture plate, placing the cell culture plate in a 28 DEG C incubator for inverted culture for 2 hours, then adding a DMEM / F-12 complete medium containing a growth additive and not containing serum, and placing the cell culture plate in the 28 DEG C incubator; when the tissue structure generates a large number of secretory vesicles, collecting the culture solution, adding a new culture solution, taking the collected culture solution as a natural hirudin crude product, and separating and purifying the natural hirudin crude product to obtain the natural hirudin. The method can achieve the effects of long-term survival in vitro and generation of a large amount of secretions.

The invention relates to recombinant pichia pastoris and application of the recombinant pichia pastoris in production of Brazilian sweet protein, and belongs to the technical field of fermentation. According to the invention, in the fermentation process of the Brazilian sweet protein, part of the Brazilian sweet protein is gathered and degraded in vacuoles, so that the Brazilian sweet protein secreted to extracellular cells is reduced, and finally, the yield of the Brazilian sweet protein in fermentation liquor is not high. Through research, the invention finds that the knockout of the protease with the amino acid sequence as shown in SEQ ID NO.2 or SEQ ID NO.3 can improve the exocytosis amount of the brazitame protein, related process parameters in the fermentation process are further optimized, the yield of the brazitame protein in a 3L fermentation tank reaches 4.01 g / L, and the high yield of the brazitame protein is realized.

The present invention relates to a composition for body fat reduction containing extracts of Taraxacum platycarpum and Lonicerae japonica as active ingredients. It has been confirmed that a mixed extract of Taraxacum platycarpum and Lonicerae japonica according to the present invention reduces body weight and body fat when administered to mice with obesity induced by a high-fat diet. Also, it has been confirmed that the mixed extract reduces the size of fat tissue, reduces fat vacuoles (pore) in liver tissue, reduces total cholesterol, triglycerides, and LDL-cholesterol in the blood, and increases HDL-cholesterol. Furthermore, it has been confirmed that the mixed extract suppresses the expression of PPAR-γ which is related to fat synthesis. In addition, it has been confirmed that the mixed extract has a synergistic effect on the regulation of intracellular fat accumulation and fat metabolism factors in cells in which fat accumulation is induced.

The present invention provides a system of polynucleotides including: (a) a first polynucleotide including a nucleic acid sequence encoding a sorting signal sequence selected from an apoplast sorting signal sequence or a vacuole sorting signal sequence and a milk protein; (b) a second polynucleotide including a nucleic acid sequence encoding a sorting signal sequence selected from an apoplast sorting signal sequence or a vacuole sorting signal sequence and a first copy of a mammalian kinase; and (c) a third polynucleotide including a nucleic acid sequence encoding a Golgi sorting signal sequence and a second copy of said mammalian kinase, wherein the first polynucleotide, the second polynucleotide, and the third polynucleotide are operably linked. Further provided are cells and compositions including the system of polynucleotides, as well as methods of suing same, such as for producing a recombinant milk protein or a plurality thereof.

The invention belongs to the field of marine natural product chemistry, and particularly relates to a marine fungus-derived alkaloid derivative and application thereof in prevention and treatment of pepper phytophthora blight. The marine fungus crude extract disclosed by the invention shows very strong antifungal activity on phytophthora capsici under the concentration of 300 mu g / mL. Seven monomeric compounds are separated and identified, NEA shows strong antifungal activity to phytophthora capsici and is stronger than positive control azoxystrobin, the NEA effectively inhibits hypha growth, sporangium formation and zoospore movement of phytophthora capsici hyphae, and meanwhile, the protective ability of pepper plants is enhanced; nEA causes obvious morphological change of phytophthora capsici protonema, so that cell membrane integrity damage, vacuole abnormal expansion, vacuole rupture, serious plasm-wall separation and disappearance of most organelles are caused. In addition, the content of MDA in the phytophthora capsici mycelium induced by the NEA is increased, the activity of antioxidant enzyme is reduced, and the level of soluble protein is reduced.

Providing a new method for determining gene mutations in JMML. [Solution] A method for determining gene mutations in juvenile myelomonocytic leukemia (JMML), comprising the step of determining that the subject has gene mutations in the CBL group or NF1 group if the distribution of monocyte morphology in a blood sample taken from the subject is as follows: (1) if there is a lot of A and a little D, the subject is in the PTPN11_somatic group; (2) if there is a lot of B, the subject is in the PTPN11_germline group; (3) if there is a lot of C, the subject is in the KRAS group; (4) if there is little A and C and a lot of B and D, the subject is in the NRAS group; or (5) if the subject does not fall into any of the above categories (1) to (4), the subject is in the CBL group or NF1 group. A is a monocyte that has cytoplasmic vacuoles and a club-shaped nucleus; B is a monocyte that does not form a cytoplasmic vacuole and has a club-shaped nucleus; C is a monocyte that has cytoplasmic vacuoles and a round nucleus; D represents a monocyte that does not form a cytoplasmic vacuole and has a round nucleus.