217 results about "Hemolysin" patented technology

The invention relates to a mutant α-hemolysin (α-HL) pore which is useful for detecting one or more nucleotides by stochastic sensing. The pore is particularly useful for sequencing DNA or RNA. A molecular adaptor that allows detection of the nucleotide(s) is covalently attached to the pore. The pore is specifically modified to facilitate positioning of the adaptor and may be modified to facilitate covalent attachment.

The invention relates to a mutant α-hemolysin (α-HL) pore which is useful for detecting one or more nucleotides by stochastic sensing. The pore is particularly useful for sequencing DNA or RNA. A molecular adaptor that allows detection of the nucleotide(s) is covalently attached to the pore. The pore is specifically modified to facilitate positioning of the adaptor and may be modified to facilitate covalent attachment.

The disclosure below provides a protein export system for efficiently producing recombinant protein from a host cell. In a preferred embodiment, the protein export system utilizes protein export machinery endogenous to the host bacterium into which the protein export system vector is introduced.

The invention relates to the technical field of human blood detection. A flow cytometry detection fluid circuit system comprises a diluent injection system, a hemolysin and blood sample injection system and a pressure system, wherein for the diluent injection system, a diluent injector is driven by a motor, and the diluent injection system is connected with a diluent barrel to suck a diluent to inject to a flowing chamber, a reaction tank and a sampling needle cleaning device; for the hemolysin and blood sample injection system, a hemolytic agent injector is driven by a motor, the hemolysin and blood sample injection system is used for sucking the hemolytic agent to inject to the reaction tank, and the sample injection sucking collects a distribution blood sample to inject to the reaction tank; for the pressure system, a diaphragm pump is connected with a pressure chamber to form positive and negative pressures, the pressure system is used for pumping waste liquid of the sampling needle cleaning device, sucking a sample in the reaction tank to a sample injection port of the flowing chamber, carrying out gas blowing stirring and evacuation on the reaction tank, and pumping the diluent to the reaction tank. According to the invention, the number of reaction tanks is reduced, system structure and complexity are simplified, and miniaturization and low cost are realized for machines.

Described herein are engineered alpha-hemolysin subunits having mutated oligomerization domains for assembling into heptameric nanopores in lipid bilayers.

The invention relates to genetically engineered bacteria for efficiently secreting, expressing and reconstructing cutinase and a method for constructing the same, belonging to the field of the bioengineering technology. The genetically engineered bacteria are escherichia coli BL21 (DE3) which carry two recombinant plasmids, and the recombinant plasmids are respectively plasmid pSTV28 carrying the specific genes in the Alpha-hemolysin A (hly A) pathway and plasmid pET20b(+) containing the cutinase-hly As genes. The method for constructing the genetically engineered bacteria for efficiently secreting, expressing and reconstructing cutinase comprises the following steps: constructing two key recombinant plasmids and transforming the constructed recombinant plasmids in the escherichia coli BL21 (DE3) to obtain the genetically engineered bacteria for efficiently secreting, expressing and reconstructing cutinase. The cutinase is produced by using the genetically engineered bacteria through culturing liquid and inducing and expressing the cutinase. The cutinase Tfu_0883 for thermophilic monospore bacteria of Thermobifida Fusca WSH03-11 is used as the report protein. The shaking flask fermentation shows that the extracellular output of the cutinase is 306U / mL which is 1.7 times of the output of the cutinase which adopts the II-type secreting pathway in the preliminary working process in the research laboratory. The cutinase is secreted and expressed efficiently.

The invention aims at providing an LAMP method for rapidly detecting the real-time turbidity Listeria monocytogenes, for making up the defects of the prior art. The method comprises the following step: firstly, providing an LAMP primer group for detecting Listeria monocytogenes, wherein the nucleotide sequences of the LAMP primer group are as shown in SEQ ID NO:1-4. A set of LAMP primers are designed aiming at the hemolysin gene of the Listeria monocytogenes, a real-time turbidity instrument is adopted to detect the Listeria monocytogenes, and the specificity and the sensitivity of the method are verified, so that a rapid, specific and sensitive Listeria monocytogenes LAMP detection method is established.

The invention discloses a kit for determining glycosylated hemoglobin and a preparation method thereof. The kit comprises three liquid components a reagent R1, reagent R2 and a reagent R3 that are independently to each other. The reagent R1 consists of: a latex glycine buffer solution, polyethylene glycol-2000, Tween-80, glycerin, bovine serum albumin, and sodium azide, the reagent R2 consists of: a glycine buffer solution, an anti-IgG antibody, an anti-HbA1c antibody, sodium chloride, bovine serum albumin and sodium azide, and the reagent R3 consists of: a hemolysin diluent. The preparation method includes: preparing the reagents according to the component content; mixing a to-be-determined sample with the reagent 3, and using the hemolysin diluents to treat the to-be-determined sample; mixing the treated to-be-determined sample with the reagent R1 and reagent R2 and carry out full reaction; employing a fully automatic biochemical analyser to determine the absorbance difference after reaction; and calculating the concentration of glycosylated hemoglobin in the sample according to an absorbance change value. The kit provided by the invention has the advantages of high accuracy, and convenient operation, etc.

The system comprises a DNA construct comprising: a) a first nucleic acid sequence containing the nucleotide sequence coding for a product of interest; b) a second nucleic acid sequence containing the nucleotide sequence coding for a dimerization domain; and c) a third nucleic acid sequence containing the nucleotide sequence coding for Escherichia coli α-hemolysin (HlyA) or for a fragment of said protein comprising the recognition signal of the E. coli hemolysin (Hly) transport system secretion mechanism. It is applicable in producing recombinant dimeric proteins.

The present invention provides a reliable method to determine lysin of ferrohemoglobin, and meanwhile white cell can be divided into two subgroups for counted separately. How to use the lisin is also included in the present invention. The lysin with no cyanide includes at least one type of surfactant group dissolve red cell sufficiently and to release magnitude of magnitude ferrohemoglobin, at least one type of positive ion surfactant, at least one type of negative ion surfactant and at least one type of non-ion surfactant.

The invention relates to an immunogenic composition, characterized in that it comprises an adenyl cyclase-hemolysin (AC-Hly) protein, or an immunogenic portion of this AC-Hly, of a strain of Bordetella chosen from B. Tertussis, B. parapertussis or B. bronchiseptica, and in that it comprises, in addition, a bacterial extract containing the expression products of the vrg genes of a strain of Bordetella chosen from B. pertussis, B. parapertussis or B. bronchiseptica, or a portion of these expression products which is sufficient to induce an immune response in a host to which the extract might be administered.

The invention relates to immunogenic compositions comprising mutant Streptococcus pneumoniae pneumolysin proteins. The invention further relates to such proteins and nucleic acids encoding these proteins. In particular embodiments, the invention is directed to an isolated mutant pneumolysin (PLY) protein, wherein the mutant PLY protein differs from the wild type PLY protein by the presence of a mutation within the region of amino acids 144 to 161 of the wild type sequence, such that the toxicity of the mutant is reduced relative to that of the wild-type protein. In particular embodiments, the mutant PLY protein differs from the wild type protein by the substitution or deletion of amino acids within this region, including the deletion of two adjacent amino acids within the region of amino acids 144 to 151 of the wild type sequence.

Described herein are variants of alpha-hemolysin having at least one amino acid substitution at H35G, E111N, M113A, and / or K147N in the mature, wild-type alpha-hemolysin amino acid sequence. In certain examples, the variant may have a substitution at E111S, M113S, T145S, K147S, or L135I in the mature alpha-hemolysin amino acid sequence. The α-hemolysin variants may also include a substitution at H144A and / or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having an increased nanopore lifetime. Further, provided are variants that, in addition to providing increased lifetime, provide a decreased time-to-thread. Hence, the variants provided herein both increase nanopore lifetime and improve efficiency and accuracy of DNA sequencing reactions using nanopores comprising the variants.

The invention discloses a method for detecting the DNA glycosylase activity by alpha-hemolysin nanopores. According to the method, a partial complementary double-chain DNA is designed and is used as aDNA glycosylase hOGG1 substrate; then, the double-chain DNA is fixed on the surface of a streptomyces affinity magnetic bead; a double-chain DNA-magnetic bead compound is formed to be used as a probe. When the DNA glycosylase hOGG1 exists, damaged basic groups 8-methoxyguanine on the double-chain DNA substrate can be specially recognized; the damaged basic groups are cut away; the DNA framework is cut away. The alpha-hemolysin nanopores are used for detecting the output DNA; the frequency of the generated blocking signals has positive correlation to the activity of the hOGG1. Therefore the method is used for detecting the DNA glycosylase activity, and has the advantages of high sensitivity, high efficiency, labeling avoidance and amplification avoidance.

The invention relates to a vibrio cholerae typing and virulence gene detection kit and a vibrio cholerae typing and virulence gene detection method. The kit comprises DNA extracting solution, PCR reaction solution, a Taq enzyme system, a positive quality control product and a negative quality control product, wherein the PCR reaction solution consists of PCR reaction buffer, four pairs of forward and reverse primers for the specificity of vibrio cholerae and four probes for the specificity of the vibrio cholerae; the primers and probes are designed according to the specificity conservative areas of nucleic acid sequences of a hemolysin gene, an O-antigen gene of O139 type vibrio cholerae, the O-antigen gene of O1 type vibrio cholerae and the virulence gene of cholera toxin (CTX). The detection kit and the detection method ensure a reliable and stable result, are easy and fast to operate and can realize the typing of the vibrio cholerae and fast judgment about whether the vibrio cholerae has a virulence gene or not, thereby facilitating the tracing and detection of the public health-related events caused by the vibrio cholerae and facilitating the routine monitoring of the vibrio cholerae.

The invention discloses a golden staphylococcus alpha-hemolysin, a polynucleotide coded as the alpha-hemolysin and a biological method of producing the alpha-hemolysin by the gene engineering technology; at the same time, the invention also discloses the purpose of the golden staphylococcus alpha-hemolysin. The golden staphylococcus alpha-hemolysin of the invention establishes a good foundation for the further research of the pathogenic and immune mechanism and the bacterin of the golden staphylococcus alpha-hemolysin.

The invention discloses compounded donkey milk powder for improving immunity. According to the formula, donkey milk powder is used as a main raw material, Chinese yam, lily, walnuts and lobed kudzuvine root, which can be eaten as both medicines and food, are added, and prebiotics with fructo-oligose and galacto-oligosaccharide, and two probiotics, namely, lactobacillus rhamnosus and lactobacillus plantarum, are additionally supplemented. By virtue of the compounded donkey milk powder, the immunity can be remarkably improved, specifically, the conversion capability of lymphocyte (cellular immune function) is remarkably improved, the production of serum hemolysin (humoral immunity function) is increased, and the carbon clearance capability (mononuclear macrophage function) and the killing activity of NK (Natural Killer) cells (NK cell activity) are enhanced.

The invention provides bifidobacterium lactis BL-99 with a function of enhancing the immunity and application thereof and belongs to the technical field of microbes. The bifidobacterium lactis BL-99 is preserved in the China General Microbiological Culture Collection Center CGMCC on April 26, 2018, has a preservation number of CGMCC No.15650, has high gastric acid tolerance and intestinal juice tolerance, can remarkably increase antibody-producing cells and half value of hemolysin HC50 and activate the activity of NK cells, can be used for preparing foods and the like with a function of enhancing the immunity, and has an extensive application prospect.

An isolated strain of Enterococcus faecalis GALT deposited under number C E C T 7121 of the group of lactic bacteria is disclosed, which is capable of surviving and colonizing the gastrointestinal tract of humans and / or animals and showing beneficial probiotic activity for the health of humans and animals. The strain E. faecalis GALT and / or a culture supernatant and / or metabolites thereof shows no in vitro multiresistance to antibiotics of common use in human clinics as glycopeptides, such as vancomycin, teicoplanine; carbapenemes, such as impipenem, meropenem; and ampicillin. The strain E. faecalis GALT contains no red blood cell-destroying hemolysins of human, ovine and equine origin; and it does not produce any gelatinase, DNase and decarboxylases. The strain E. faecalis GALT is useful for the preparation of a composition intended for the treatment and / or prophylaxis of disorders associated with colonization by pathogenic microorganisms of the gastrointestinal tract; for use as a regulator of the immune response in human and animals, as well as for the preparation of a composition. The invention is also directed to methods and uses of the strain E. faecalis GALT.

Vital mononucleosis listeria RT-PCR fast inspecting reagent kit and its method are disclosed. The process is carried out by taking mRNA of mononucleosis listeriolysin gene hlyA as target, extracting sample RNA, RT-PCR amplification reacting and electrophoresis inspecting. It is fast, has better sensitivity and operability. It can be used in environmental inspection, food production and commodity quarantine.

The invention discloses a methicillin-resistant staphylococcus aureus (MRSA) vaccine recombinant protein antigen HI2, a preparation method and application thereof. The recombinant protein comprises a staphylococcus aureus (alpha) hemolysin (Hla) fragment and a ferrum surface determination protein (IsdB) fragment, wherein the Hla and the IsdB fragments are fused by a connecting peptide. The protein can be applied to preparation of a subunit vaccine and relative detection kits for resisting infection from methicillin-resistant staphylococcus aureus. According to the preparation method, the fused protein can be subjected to recombinant expression through the genetic engineering technology, therefore high expression quantity is gained, the separation and purification are convenient to carry out, and high efficiency and safety are ensured. As the animal experiment shown, the antigen can effectively stimulate an organism to perform higher humoral immune response and excellent immunoprotection.

The invention relates to an echinacea extract product, a preparation method and an application, the extract product comprises the following components by weight percentage: content of phenolic compounds is higher than 12%, content of Echinacea polysaccharide is higher than 20%, and chicoric acid content is higher than 4%. The extract product has obvious promotion effect to the generation of mice hemolysin antibody, the echinacea extract product can enhance humoral immunity, and the effect of the echinacea extract product is better than that of the prior art.

The invention discloses a detection method of human peripheral blood lymphocytes. The preparation method comprises the following steps of: taking human anticoagulation peripheral blood; adding anti-human CD3, CD4, CD8 and CD45 monoclonal antibodies and anti-human CD3, CD56, CD16 and CD45 monoclonal antibodies, then adding a hemolysin working solution containing a fluorescent probe to respectivelyobtain the percentage and absolute count value of a human peripheral blood T lymphocyte subset and an NK lymphocyte subset and achieve the detection of human peripheral blood lymphocytes. The method has the advantages of simplicity and convenience in operation, long sample stable preservation time, high accuracy, multi-index output and the like.

The invention discloses a method for detecting vibrio parahemolyticus through combination of unlabelled fluorescent PCR and HRMA, which is characterized by including the following steps: firstly, a primer pair is designed according to a TLH (thermolabile hemolysin) gene of the vibrio parahemolyticus; secondly, after DNA (Deoxyribonucleic acid) samples are extracted, unlabelled fluorescent PCR amplification is performed by utilizing the designed primer pair; and thirdly, an amplification curve of a PCR amplification product and the HRMA are analyzed by application software. The purposes of the method are to overcome defects of the prior art and provide a method, which is simple, convenient and fast to operate, accurate in detection result and low in usage cost, for detecting the vibrio parahemolyticus through the combination of the unlabelled fluorescent PCR and the HRMA.

The invention belongs to the field of molecular biology, and specifically relates to a detection primer for rcnobcterium pyogenes and a detection kit. A specific and sensitive PCR method is established for a hemolysin PLO gene sequence of rcnobcterium pyogenes, so that the rcnobcterium pyogenes can be accurately detected, and other pathogens for goat pyogenic infection are distinguished. Positivecontrol bacteria and negative control bacteria are provided for a user to determine the quality of the kit and the accuracy of an assessment method. The kit and the method provided by the invention can be used in fundamental conventional animal epidemic disease diagnose laboratories, target stripes are clear and easy to distinguish, a PCR reaction can be completed within one hour, and the kit andthe method are significant for monitoring the reproduction of bacterium, occurrence and propagation of diseases and timely prevention and curing of diseases.

The invention relates to protein peptide powder for enhancing immunity and a preparation method and application thereof. The protein peptide powder is prepared from the following raw materials by weight: 30%-45% of marine fish skin collagen oligopeptide powder, 30%-50% of soybean oligopeptide powder, 10%-20% of Isomaltooligosaccharide, 8%-13% of polydextrose and 0%-0.5% of a sweetener. The protein peptide powder can increase macrophage phagocytic rate and phagocytic index and serum hemolysin content, enhances lymphocyte proliferation and NK cell activity, promotes antibody formation cell number increase, improves the body's immunity, and enhances body immunity.

The invention belongs to the field of molecular biology and immunology. The invention relates to three truncated proteins of a staphylococcus aureus iron regulation surface determining protein B (IsdB) and a staphylococcus aureus alpha-hemolysin (Hla) attenuated protein, and their coding genes, preparation method and use. The invention also relates to the combined use of the three truncated proteins and the alpha-hemolysin attenuated body.

The invention provides a quick detection method and a fluorescent PCR (polymerase chain reaction) detection kit invented according to the typical biochemical characteristics of sucrose, arabinose, mannose, oxidase, haircuts, salt-free peptone water and the like of the vibrio cholerae. The invention designs primers and TaqMan probes according to the outer-membrane protein gene (ompW), the toxin expression regulation protein gene (toxR) and the hemolysin gene (hlyA), wherein the vibrio cholerae outer-membrane protein gene is used as a characteristic gene for identifying the vibrio cholerae, andthe toxin expression regulation protein gene and the hemolysin gene are used for discriminating toxin genes carried by the vibrio cholerae. The fluorescent detection method for detecting the vibrio cholerae and related toxin genes thereof is provided based on the three genes. According to the detection method, strains are primarily screened through biochemical reaction, the strains with positive results are confirmed by fluorescent quantitative PCR, and the strains with positive ompW genes are vibrio cholerae. The vibrio cholerae and related toxin genes thereof can be quickly, accurately and specifically detected by the biochemical reaction and fluorescent quantitative PCR.