63 results about "Bacterial lysis" patented technology

The present invention provides bacterial vectors and fusion proteins containing a TTSS polypeptide, compositions of such fusion proteins including polynucleotides, and methods of delivering one or more genes into a target cell that involve contacting the cell with a composition that includes such a fusion protein. Compositions and methods of gene delivery that involve a bacterium and a TAT, Antp, or HSV VP22 polypeptide are also disclosed. The invention also concerns methods of delivering one or more genes into a target cell utilizing a bacterium capable of becoming internalized within the cell, wherein the bacterium includes one or more genes targeted for delivery to the cell, a gene encoding an RNA polymerase, and a gene that causes lysis of the bacterium.

The invention provides methods and compositions for selectively isolating total bacterial mRNA. The general method comprises contacting a bacterial lysate comprising total bacterial mRNA and nonisolated bacterial polysomes with an exogenous enzyme under conditions wherein the enzyme selectively modifies the mRNA to form modified mRNA, and isolating the modified mRNA. In particular embodiments, the enzyme is selected from a poly(A) polymerase, a RNA ligase and a terminal deoxynucleotidyl transferase. Depending on the enzyme, the modified mRNA may have any of a variety of modifications, such as a 3' tail, particularly a poly(A) tail, a specific sequence tag, a detectably labeled nucleotide, etc.

Disclosed herein are compositions and methods for regulating redox status and/or reducing oxidative stress in a subject, the methods and compositions comprising TLR agonists comprising bacterial lysates and/or lysate fractions. Also disclosed are compositions and methods comprising bacterial lysates and/or lysate fractions formulated or administered in combination with one or more other therapeutic or pharmaceutical agents.

Chimeric anti-microbial proteins, compositions, and methods for the therapeutic and prophylactic treatment of plant diseases caused by the bacterial pathogen Xylella fastidiosa are provided. The anti-microbial proteins of the invention generally comprise a surface recognition domain polypeptide, capable of binding to a bacterial membrane component, fused to a bacterial lysis domain polypeptide, capable of affecting lysis or rupture of the bacterial membrane, typically via a fused polypeptide linker. In particular, methods and compositions for the treatment or prevention of Pierce's disease of grapevines are provided. Methods for the generation of transgenic Vitus vinefera plants expressing xylem-secreted anti-microbial chimeras are also provided.

The present invention discloses a method for synthesizing ferulic acid through an enzyme method, wherein the methyl on S-adenosyl methionine (SAM) is transferred onto caffeic acid so as to finally form the product ferulic acid. The method comprises: a) highly expressing ligusticum chuanxiong hort caffeic acid-O methyltransferase(COMT) through fermentation of escherichia coli, collecting bacteria through centrifugation, re-suspending the bacteria with a Tris-HCl buffer solution, and carrying out ultrasonic bacterial lysis to obtain a mixed enzyme solution containing the ligusticum chuanxiong hort COMT; and b) transferring the methyl provided by SAM to the caffeic acid through the ligusticum chuanxiong hort COMT obtained in the step a), and finally converting into the ferulic acid. The method of the present invention has advantages of simple process, mild reaction condition, high conversion efficiency, low conversion cost, and the like.

A method of effecting lysis of acid-fast bacteria, comprising heating acid-fast bacteria in a liquid containing a non-ionic surfactant at a temperature of below the boiling point of the liquid. This method enables accomplishing secure lysis of acid-fast bacteria in a simple manner within a short period of time without the use of special apparatus and agent and enables extracting genes. The heating is preferably conducted at 96° C. for 10 min. As the nonionic surfactant, use can be made of a d-sorbitol fatty acid ester, a polyoxyethylene glycol sorbitan alkyl ester, a polyoxyethylene glycol p-t-octylphenyl ether or the like. The pH value of the liquid is preferably 8, and the liquid preferably contains EDTA. It is also preferred that before the heating, the acid-fast bacteria be treated with lipase.

The invention relates to a method and device for continuously removing cell/bacterial lysis floccules. The method comprises the following steps of: 1) arranging a solid-liquid separation device, wherein the solid-liquid separation device comprises a motor and a roller driven by a transmission mechanism, the roller is internally provided with blades extending forwards in a spiral shape, a screen is arranged outside the roller, a collection bed is arranged below the roller, and the liquid outlet of the collection bed is connected with a downstream deep bed filter by virtue of a liquid pump; 2) starting a motor to drive the roller to rotate, feeding a lysis solution containing solid floccules into the roller while the lysis solution spirally advances under the driving of the blades, and carrying out solid-liquid separation; and 3) causing a liquid to flow into the collection bed by virtue of the screen, and taking the solid floccules out of the roller by virtue of the blades. In the invention, the lysis solution is advanced under the actions of the blades and the screen while the solid-liquid separation is carried out, the liquid flows into the collection bed by virtue of the roller, and dehydrated solid floccules can be discharged from a solid collection hole, thus the method and device provided by the invention can be widely applied to a production process for continuously removing cell/bacterial lysis floccules in a large scale.

The invention discloses application of pyroptosis-associated protein GSDMD (Gasdermin-D) for preparing a bacterial ghost vaccine. A bacteriolysis plasmid containing a pyroptosis-associated protein GSDMD coding gene is constructed and is converted into salmonella enteritidis; under the induction of gum sugar, bacteria are split to successfully obtain a salmonella novel bacterial ghost vaccine. An experiment proves that GSDMD mediated bacteriolysis has an extremely long persistent period, and a splitting rate is 99.9985% or more. After a novel bacterial ghost immune mouse prepared by the invention is adopted, an organism can be stimulated to generate powerful humoral immunity and cellular immunity; protective immunity response is induced; an experiment animal can be protected so as to resistsalmonella infection. The salmonella bacterial ghost vaccine prepared by the invention has a good immune protection effect.

The invention discloses a special bacterial genome DNA extraction kit and method for septicemia. The kit consists of a buffer solution GPA, a buffer solution GRB, a bacterial lysis solution DA, a bacterial lysis solution DB, a buffer solution DC, a washing solution PE, an eluent EB and protease K. According to the extraction kit and method provided by the invention, red/white cells are separated by virtue of the buffer solution GRB, so that the influence and interference of human genome can be removed to the greatest extent; bacterial cells are cracked and broken by virtue of SDS and the protease K, and protein is degraded, so that bacterial genome DNA is released; and by virtue of a silica gel column separation and purification method, the bacterial genome DNA with high quality can be obtained.

The method for isolating nucleic acids from a food sample comprising the following steps: a) obtaining a food enrichment culture; b) transferring a portion of the food enrichment culture into a reaction vessel thereby providing a food enrichment sample and providing a water-immiscible phase in contact with the food enrichment culture; c) lysing the food enrichment sample to provide a lysed sample; d) isolating nucleic acids from the lysed sample. Food enrichment culture samples are known to contain high concentrations of organic and/or liposoluble inhibitors. By contacting the enrichment culture sample with a water-immiscible phase before the actual DNA extraction procedure starts, part of the lipophilic inhibitors is expected to cross the phase interface due to an enhanced solubility in the organic phase and thereby become depleted. The water-immiscible phase provided according to the invention interacts directly with the sample material throughout bacteria lysis and DNA extraction thereby optimizing the subsequent DNA purification processes due to the depletion of food-borne inhibitors. This method yields nucleic acids, in particular DNA that is of an improved quality and purity to be used in a subsequent PCR reaction to detect pathogen nucleic acids in the isolated nucleic acids.

The invention, which belongs to the technical field of bacterial lysis devices, discloses a portable bacterial lysis device based on a centrifugal microfluidic technology and a using method thereof. The device comprises upper and lower circular plates, a disc micro-fluidic chip, and a rotating shaft, wherein the disc micro-fluidic chip and the rotating shaft are parallel to each other between theupper and lower circular plates. The rotating shaft passes through the parts, away from the circle centers, of the circulate plates but is not in contact with the circular plates; and the rotating shaft passes through the circle center of the disc micro-fluidic chip. A plurality of magnets are respectively arranged on the circular plates and are uniformly distributed on each circular plate, wherein the magnets are mutually staggered in a vertical position. A plurality of bacteria lysis pools are arranged on the disc micro-fluidic chip. According to the invention, the bacterial lysis efficiencyand the sample treatment flux can be effectively improved; and other operation units can be integrated to realize real-time on-site detection of bacteria. The designed bacterial lysis device is portable, low in cost, easy and convenient to operate; the operation is automatic; and the device is used flexibly. The portable bacterial lysis device can meet different requirements and has the broad application prospects.

The invention discloses an alpha-1, 3 galactosyl transferase fusion protein and a preparation method thereof. The alpha-1, 3 galactosyl transferase fusion protein provided by the invention comprises TRX (thioredoxin) and catalyzes the formation of an alpha-galactose epitope. The invention also provides a bacterium for generating the alpha-1, 3 galactosyl transferase fusion protein. The fusion protein provided by the invention completely has the activity of alpha-1, 3 galactosyl transferase, is expressed in a soluble way, further, is higher in expression quantity, occupies above 20% of a soluble bacterial crackate, and has a favorable industrial application prospect.

The invention discloses a method for quickly extracting a genome DNA (Deoxyribo-Nucleic Acid) of riemerella anatipestifer, and a kit. The method comprises the following steps: 1) lysing bacteria; 2) precipitating bacterial proteins; 3) removing the bacterial proteins; 4) precipitating the genome DNA; 5) deionizing; and 6) dissolving the DNA, wherein a bacterial lysis buffer solution used in the step 1) contains 38-42 mM of Tris-acetate having the pH of 7.8, 18-22 mM of sodium-acetate, 0.8-1.2 mM of EDTA (Ethylene Diamine Tetraacetic Acid) and 0.8-1.2% of SDS (Sodium Dodecyl Sulfonate). The extraction method provided by the invention is high in extraction efficiency, and the genome DNA of riemerella anatipestifer extracted by the method is high in purity, which lays a strong foundation for subsequent molecular biologicalstudy; and the extraction method is simple and feasible to operate, and high in practicability.

The invention relates to a method for preparing lactoferricin and a method for applying the lactoferricin in bacterial inhibition of foods. The method for preparing the lactoferricin comprises the following steps of: (i) activating genetic engineering bacteria; (ii) performing bacterial lysis; (iii) separating an inclusion body and performing lysis; (iv) extracting fusion protein and performing acid hydrolysis; and (v) separating by using a chromatographic column. The method is feasible, easy to operate and low in cost; and the lactoferricin prepared by the method can be used for bacterial inhibition of aquatic products. The method for applying the lactoferricin comprises the following steps of: cleaning an aquatic product, and soaking in a solution of the lactoferricin at normal temperature or at the temperature of between 0 and 4 DEG C for 0.5 to 10 hours.

The invention relates to bacterial genome extraction, in particular to a method of extracting genome DNA from methicillin-resistant staphylococcus aureus (MRSA). Lysozyme, glycyl-glycine endopeptidase and nuclear cracking liquid are utilized for cracking of MRSA. The method includes following steps: (1), performing bacterial culture: inoculating MRSA to a liquid culture medium, and performing shaking culture; (2), performing bacterial collection: centrifuging MRSA overnight culture liquid to remove supernate, adding EDTA for resuspended precipitation; (3), performing bacterial cracking: sequentially adding lysozyme and glycyl-glycine endopeptidase into a resuspension for incubation, centrifuging to remove supernate after incubation is completed, and performing nuclear cracking; (4), removing RNA and protein; (5), precipitating and washing. Genome DNA extracted by the method is high in purity and concentration, requirements on identification and sequencing of specific genes can be met, and the method is time-saving, efficient and simple to operate.