72 results about "Deoxyribonuclease" patented technology

The present invention is separating and culturing process of human amnion mesenchyme stem cell and its medical composition. The separating and culturing process includes digesting human amnion successively with trypsin, collagenase and deoxyribonuclease, and filtering to prepare single cell suspension; culturing in DMEM / F12 culture medium with VDMEM and VF12 in the equal ratio and containing ox embryo blood serum in 10-20 vol% and basic fibroblast growth factor of ultimate concentration 10-20 ng / ml inside a culture box at 37 deg.c, saturated humidity and CO2 in 5 vol%; and replacing liquid and culture passage to proliferate and purify human amnion mesenchyme stem cell. The process has wide material source no ethnic limitation and wide application foreground. The medical composition may be used in various kinds of treatment.

The invention relates to compounds, compositions and methods for biofilm disruption and prevention. In particular, the invention relates to pharmaceutical compositions for the disruption of biofilm and prevention of biofilm in patients. The invention also relates to anti-biofouling compositions for the disruption of biofilm and prevention of biofilm on surfaces. The invention also relates to the removal of biological material from surfaces. The compositions of the invention include microbial deoxyribonucleases.

Methods and compositions for inhibiting and / or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease. The invention also relates to kits for the performance of various microbiological procedures, which kits comprise the nuclease inhibitors described herein.

The invention discloses a preparation method of a natural-tissue-derived decellularized and decalcified bone material. According to the method, any bone tissue of a mammal is treated with a protease inhibitor-containing normal saline buffer, an organic solvent, Tirton X-containing PBS (phosphate buffer saline), SDS (sodium dodecyl sulfonate)-containing PBS, pancreatin-containing PBS, deoxyribonuclease-containing PBS, an EDTA (ethylene diamine tetraacetic acid) isotonic solution and ultrasonic waves, and the decellularized and decalcified bone material is obtained. Cancellous bones and cortical bones can be decellularized completely and simultaneously, the conditions are mild, damage to an ECM (extracellular matrix) is avoided, rapidness and stability are realized, and the obtained decellularized and decalcified bone material has the advantages of good biocompatibility, high plasticity, high biomechanical strength and the like and can be used for clinically repairing bone regeneration and repair disorder such as bone defects, nonunion and the like caused by various causes.

The invention relates to a Pb<2+> detection chip based on deoxyribonuclease as well as making and application methods. The invention is characterized in that a 17DS substrate chain of a 17E enzyme chain of deoxyribonuclease 8-17 with strong specificity response is cut off, so that part or all of the cut-off 17DS substrate chain falls off. Since the 17DS substrate chain is previously labeled by fluorescence, the cut-off operation weakens the fluorescence signal. The application method is characterized by comprising the following steps: when the lead ion detection chip is used for lead ion detection, putting the sample to be detected on the chip, holding for a period of time, scanning the chip by using a chip signal analysis system, and analyzing the fluorescence signal. The Pb<2+> is detected through the variation of the fluorescence signal; and the fluorescence signal gets weakened as the concentration of Pb<2+> in the sample gets higher. The detection range of the concentration of Pb<2+> is 1nM-10mu M.

The present invention relates to antimicrobial deoxyribonuclease-based compositions that inhibit growth and proliferation of Staphylococcus aureus bacteria. The present invention also relates to methods of administering the compositions in the treatment and prevention of S. aureus infections. The present invention also relates to methods of administering the compositions in the eradication of S. aureus nasal carriage, in order to prevent the transmission of S. aureus bacteria

The invention discloses a method for safely and quickly extracting a genomic deoxyribose nucleic acid (DNA) from blood. The method comprises the following steps of: lysing a blood cell by using TritonX-100 and sodium dodecyl sulfonate (SDS), precipitating a protein with saturated NaCl, separating the protein through centrifugation, and reducing the mechanical shearing damage of a solution to the DNA; preventing and inhibiting the degradation of deoxyribonuclease (DNase) to the DNA; excluding the pollution caused by organic solvent, metal ions and other cnucleic acid molecules; and reducing the protein, polyoses, lipids and the like to the lowest degree, wherein the extracted DNA has certain length, and substances which inhibit enzymes exist in the genomic DNA after purification. The method for extracting the genomic DNA from the animal blood is harmless, quick and simple in operation.

The invention discloses a bacillus subtilis polygene editing and expression adjusting and controlling system based on CRISPR Cpf1. Through a constructed Cpf1 expression vector pHT-XCR6 and a constructed crRNA array expression vector pcrF11, complete knockout of 2 genes, basic group modification of 6 genes and knock-in of 1 gene can be completed at one time. The vector pHT-XCR6 contains NgAgo protein, and is used for promoting recA mediated homologous recombination. Besides, a vector pLCg6-dCpf1-remA (used for integral expression of a Cpf1 mutant dCpf1 inactivated by deoxyribonuclease and fusion protein of an activating transcription factor remA on a bacillus subtilis genome) and a vector pcra3 (used for integral expression of a crRNA array on the bacillus subtilis genome) are constructed,and can be used for performing transcription inhibition and activation on different genes. The invention further establishes an economic efficient crRNA array assembling method of which the name is SOMACA (Synthetic Oligos Mediated Assembly of crRNA Array), and through synthesizing short-single-strand DNA (60nt ), a needed crRNA array is inserted in the vector pcrF11 or pcra3 and is used for guiding Cpf1 or dCpf1-remA for gene editing and expression adjustment and control.

The invention discloses a staphylococcus aureus multiple PCR-MIX rapid detection kit and a detection method thereof; the detection kit comprises PCR MIX; wherein, the PCR MIX contains 2*PCR buffer, 1.0-3.0mmol / L of MgCl2, 180-220mu mol / L of dNTP each, heat-resisting deoxyribonuclease nuc gene primer of which the base sequence is SEQ NO.1 and the concentration of SEQ NO.1 is 100-600nmol / L, plasma-coagulase clfA gene primer of which the base sequence is SEQ NO.3 and the concentration of SEQ NO.4 is 100-600nmol / L, SmaI restriction fragment specific sequence primer of which the base sequence is SEQ NO.5 and the concentration of SEQ NO.6 is 100-600nmol / L, 20-60U of Taq enzyme and bromophenyl blue dye. The multiple PCR-MIX rapid detection kit and the detection method provided by the invention have simple configuration, the detection kit and the detection method can be used in industrialized production; the detection method is sensitive and has short detection cycle and strong maneuverability, so that the method can be widely applied in the fields of food hygiene, environmental monitoring, etc.

The invention discloses a method for removing cells from a human amnion. The method is characterized by comprising the following steps of: obtaining a clean semitransparent amnion, rinsing the semitransparent amnion by using a PBS (Phosphate Buffer Solution), soaking the semitransparent amnion in a comprehensive PBS liquid, shocking, rinsing the amnion by using the PBS after the amnion is taken out, successively soaking the amnion in steapsin / PBS liquid and deoxyribonuclease / PBS liquid for treatment, rinsing the amnion by using double-antibody / PBS liquid, taking out the decellularized amnion and putting the decellularized amnion in the PBS for preservation and standby. The method has the advantages that the cells are removed from the amnion in a manner that a surfactant is combined with steapsin and deoxyribonuclease, the cells can be prompted to be completely and efficiently removed from the amnion due to the combined use of the steapsin and the deoxyribonuclease, and meanwhile, the extracellular matrix is reserved to the maximum and is enabled to be free from damage.

Methods and compositions for inhibiting and / or inactivating nucleases by employing nuclease inhibitors are provided. The nuclease inhibitors comprise anti-nuclease antibodies and non-antibody nuclease inhibitors. The anti-nuclease antibodies of the present invention may be a polyclonal or monoclonal antibodies, and may be anti-ribonuclease antibodies, anti-deoxyribonuclease antibodies, or antibodies to non-specific nucleases. A preferred embodiment comprises at least two nuclease inhibitors, and is referred to as a nuclease inhibitor cocktail. In some specific embodiments, the invention concerns methods of performing in vitro translation comprising obtaining a first nuclease inhibitor, which inhibitor is further defined as an anti-nuclease antibody, and placing the anti-nuclease antibody in an in vitro translation reaction. In many cases, the in vitro translation reaction comprises at least one nuclease, which may be a ribonuclease, a deoxyribonuclease, or a nonspecific nuclease. The invention also relates to kits for the performance of various microbiological procedures, which kits comprise the nuclease inhibitors described herein.

The invention relates to compounds, compositions and methods for biofilm disruption and prevention. In particular, the invention relates to pharmaceutical compositions for the disruption of biofilm and prevention of biofilm in patients. The invention also relates to anti-biofouling compositions for the disruption of biofilm and prevention of biofilm on surfaces. The invention also relates to the removal of biological material from surfaces. The compositions of the invention include microbial deoxyribonucleases.

The present invention concerns the use of a polypeptide having deoxyribonuclease (DNase) activity for preventing, reducing or removing static electricity from an item; a composition comprising such polypeptide; a method for reducing or removing static electricity from an item; and a wipe for such use.

The invention discloses a decellularized fiber ring matrix preparation method and belongs to the technical field of animal cells or tissues. The decellularized fiber ring matrix preparation method includes taking spinal fiber ring of a healthy adult pig, sufficiently rinsing the spinal fiber ring by sterile saline solution; placing the spinal fiber ring into Tris buffer solution of concentration 10mM and with 0.05-0.5% of EDTA (ethylene diamine tetraacetic acid) and 5-50KIU / ml of aprotinin and oscillating at the temperature of 4 DEG C; then placing the spinal fiber ring into the Tris buffer solution containing 1-5% of TritonX-100, oscillating at room temperature; placing the spinal fiber ring into Tris buffer solution containing 0.05-0.5mg / ml of deoxyribonuclease, 5-50microgram / ml of ribonuclease A, and oscillating; and finally, washing by sterile saline solution with oscillating, and storing in sterile PBS (phosphate buffer solution) at the temperature of 4 DEG C. The decellurlarized fiber ring matrix preparation method is simple in preparation process, thorough in decellurlarization and complete in reserved structure, similar in matrix content and mechanical performance as those of natural fibers, and is a good tissue engineering fiber ring support material.

The invention relates to an isolated culture method of piglet myocardial fibroblasts and belongs to the technical field of cell culture in modern biotechnology. The method includes: 1, collecting cardiac tissues from a piglet 1 to 3 days old, shearing ventricular tissues, performing PBS (phosphate buffer saline) pre-cooling, and rinsing several times; 2, digesting the myocardial tissues, namely performing multiple digestion at 37 DEG C with a mixture of trypsin 0.25% and collagenase II 0.1% having a ratio of 1:1, adding DNA deoxyribonuclease (0.02 mg / mL) to reduce cell suspension viscosity and increase cell yield, and adding red blood cell lysis buffer to reduce the amount of red cells; 3, culturing piglet myocardial fibroblasts, namely re-suspending cells with DMEM (Dulbecco modified Eagle medium) high-glucose medium containing fetal calf serum 10-15%, and performing differential attachment to obtain the myocardial fibroblasts. The method is simple and easy to master, time efficient and high in success rate, the obtained piglet myocardial fibroblasts are good in form, good in activity and abundant. Certain basis is provided for establishing a cell model of piglet myocardial fibroblasts to study related diseases such as cardiac hypertrophy in future.

The invention relates to a DNA pulldown method and a kit. The method includes the following steps of pretreating magnetic beads, wherein the magnetic beads marked by BSA(bovine serum albumin) and oligonucleotide random primer closed streptavidin are used; preparing a probe-magnetic bead compound; extracting and pretreating cell nucleus protein, wherein deoxyribonuclease is added into an extracting nucleoprotein sample for incubation, the magnetic beads are added for incubation and then removed, and supernatant is collected; carrying out pulldown; collecting the protein sample. The DNA pulldown method has the following advantages that BSA and an oligonucleotide random primer are used for closing the magnetic beads in advance, and nonspecific binding between the magnetic beads, protein and genomic DNA is reduced. Nuclease contamination is prevented through systemic nuclease protection, stability and reliability of experiments are improved, repeatability is high, and the experiment process is simplified.

The present invention belongs to the field of animal leather product for medicine. The method for preparing radiation sterilized allogenous acellular dermal matrix mainly comprises the following steps: preparing skin; cutting skin with machine for separating corium and epidermis and removing cell: first removing: cleaning with pure water after immersing the skin in 10-20% of parenzyme for 30-50 minutes in 36-40 DEG C; and second removing: alternately immersing the skin piece which has a water content of 10-30% after water logging in 10-20% of deoxyribonuclease and 10-301051632074f ribalgilase for 2-4 times in 36-40 DEG C, and immersing the skin piece in 0.5-1.5% of deoxyribonuclease or ribalgilase for 5-30 minutes in 36-40 DEG C for removing the substance which can initiate the recognition reaction of host cell in the corium; immersing the processed material in common fixing agent for 50-120 minutes; and radiating for sterilization, namely executing gamma-ray radiation sterilization through electron accelerator or Co-60.

The invention relates to a target protein-induced deoxyribonuclease (DNase) cycle generation-based homogeneous immunoassay method. A detection solution of the immunoassay method is prepared from an antibody 1-DNA1 conjugate, an antibody 2-DNA2 conjugate, hemin, exonuclease III and DNA3 / DNA 4 double-stranded DNA. When a target protein exists, the target protein can be immunologically recognized by the antibody 1-DNA1 conjugate and the antibody 2-DNA2 conjugate at the same time; based on an ortho effect, a complex is formed by hybridizing DNA1 and DNA2 and then has a chain substitution reaction with DNA3 on the double-stranded DNA, so that DNA4 rich in a G4 sequence is released and combines with the hemin so as to form G-quadruple chain / hemin DNase; in addition, the DNA3 reacting with the DNA1 and the DNA2 can be recognized and cut by the exonuclease III, so that the DNA1 and the DNA2 can undergo a chain substitution reaction with another DNA3, and another DNA4 is accordingly released to produce DNase; therefore, if repeating in this way, a large amount of DNase can be produced. The DNase can catalyze the color development of tetramethylbenzidine (TMB) and the luminescence of luminol, and can be used for carrying out sensitive quantitative analysis on the target protein by means of color comparisons and chemiluminiscence. The immunoassay method is simple to operate, rapid, sensitive and high in universality, and has a certain clinical application value.

The invention relates to compounds, compositions and methods for biofilm disruption and prevention. In particular, the invention relates to pharmaceutical compositions for the disruption of biofilm and prevention of biofilm in patients. The invention also relates to anti-biofouling compositions for the disruption of biofilm and prevention of biofilm on surfaces. The invention also relates to the removal of biological material from surfaces. The compositions of the invention include microbial deoxyribonucleases.

The invention discloses a vaginal substrate material and a preparation method thereof, wherein a vaginal tissue mucous layer of a pig or a cow is taken as a carrier, and is subjected to the following treatment: a, carrying out shocking treatment by adopting deionized water solution containing phenylmethylsulfonyl fluoride or Tris solution; b, carrying out shocking treatment in a water solution containing lauryl sodium sulfate or Tris-HCl buffer solution; c, carrying out shocking treatment by adopting Tris-HCl buffer solution containing Triton X-100; d, carrying out shocking treatment in Tris-HCl buffer solution containing deoxyribonuclease and ribonuclease; and e, carrying out disinfection and freeze-drying. The vaginal substrate material provided by the invention has the biological activity and the biological mechanical property more suitable for pelvic floor repairing or vagina reconstruction.

The invention relates to a composite enzyme apophlegmatisant and belongs to the technical field of medicament. The apophlegmatisant is a solid preparation or a liquid preparation which is prepared from combination of one or more deoxyribonuclease and one or more protease and a medicinal auxiliary material; in a solid preparation prescription, the mass proportion (W / W) of the deoxyribonuclease, the protease to the medicinal auxiliary material is 1 to (0.1-1) to (50-2,000); and in a liquid preparation prescription, the mass proportion (W / W, excluding water, a propellent and other cosolvents) of the deoxyribonuclease, the protease to the medicinal auxiliary material is 1 to (0.1-1) to (0.01-100). The composite enzyme apophlegmatisant can effectively remove sputum, especially thick sputum in oral cavity, laryngeal and trachea so as to overcome the block of the sputum to respiration.

The invention relates to the field of enzyme engineering, and discloses a kit for separating single cells and applications thereof. The kit comprises VIII type collagenase, neutral protease, a trypsininhibitor, and deoxyribonuclease; and the enzyme active unit ratio of VIII type collagenase: neutral protease: a trypsin inhibitor: deoxyribonuclease is 50-90: 0.5-1: 2000-5000: 1. The invention alsodiscloses an application of the kit on separating single cells from a pancreas sample. The provided kit can extract single cells (in particular from pancreas tissues), the yield is high, high activity of single cells is maintained, and the problems that the yield of single cells extracted from pancreas tissues is low and the activity of the single cells is low in the prior art are solved.

The invention belongs to the technical field of a DNA sensor and specifically relates to a method for testing Pb2+ concentration by using the DNA sensor based on G-quadruplex. According to the invention, Pb2+ is detected only through a 'mixing-testing' step and the sensitivity and the selectivity are ultrahigh; compared with metallic ion auxiliary enzyme and RNA cracking deoxyribonuclease, the G-quadruplex DNA is more stable and no complex preparation process is required; compared with the scheme utilizing various shearing enzymes, the scheme is more economic and convenient; PPIX used as a fluorescent signal indicating unit is low-cost and has effective fluorescent characteristics; these mark-free characteristics can thoroughly reduce the test cost.

The invention relates to the field of single cell separation, and discloses a method for separating single cells from tissue. The method comprises the steps that a tissue sample is mixed with digestive juice for digestion, wherein the digestive juice contains VIII-type collagenase, neutral protease, a trypsin inhibitor and deoxyribonuclease; the ratio between enzyme activity units of the VIII-typecollagenase, the neutral protease, the trypsin inhibitor and the deoxyribonuclease is (50-90):(0.5-1):(2000-5000):1. The method can achieve high-yield and high-activity extraction of the single cellsunder the condition that the total digestion time is short, especially pancreatic tissue, and the problem that existing pancreatic tissue single cells are relatively low in yield and activity is solved.

The invention relates to a culture and cryopreservation method of amniotic mesenchymal stem cells. The culture method comprises amniotic tissue separation, amniotic mesenchymal stem cell separation, P0 generation amniotic mesenchymal stem cell culture and amplification culture. According to the culture method of the amniotic mesenchymal stem cells, at a separating stage of the amniotic mesenchymalstem cells, a special mixed enzyme digestive juice system (the final concentrations of components in the digestive juice are as follows: 1.5-2U / mL of neutral protease, 0.5mg / mL of deoxyribonuclease Iand 1mg / mL of collagenase IV) is adopted for digestion, so that the amniotic mesenchymal stem cells can be more effectively separated from amniotic tissues, and then the yield of P0 generation cellsis obviously improved. Compared with conventional two-step digestion, the culture method provided by the invention has the advantages that a one-step digestion method is adopted, operation is more convenient, and the cultured amniotic mesenchymal stem cells have good activity, high yield, high purity and good repeatability.

The invention provides a DNA probe, a reagent kit and a method for detecting deoxyribonuclease. The hairpin-shaped DNA probe serves as a substrate of deoxyribonuclease, and a fluorescence group and a corresponding fluorescence quenching group are marked at the two ends or the interior of the probe respectively. When the structure of the probe is complete, the fluorescence group and the fluorescence quenching group are located on the same DNA molecule, excited fluorescence of the fluorescence group is absorbed by the fluorescence quenching group, and the fluorescence group does not emit fluorescence; when the probe is degraded by deoxyribonuclease, the fluorescence group and the fluorescence quenching group are separated, and the fluorescence group can emit fluorescence normally. Whether deoxyribonuclease exists in a reaction system or not can be judged as long as the intensity of fluorescence emitted from the reaction system is detected with a real-time quantitative PCR instrument. The method can be widely applied to quality detection on residues of deoxyribonuclease in biological products or quantitative detection of deoxyribonuclease.

The present invention concerns the use of a polypeptide having deoxyribonuclease (DNase) activity for preventing or reducing redeposition of soil on an item during a subsequent cleaning or laundering process. The invention further concerns a detergent composition comprising a polypeptide having deoxyribonuclease (DNase) activity and a method for preventing or reducing redeposition of soil on an item during a subsequent cleaning or laundering process.