283 results about "Antibiotic resistance genes" patented technology

The invention is related to a transducing particle that comprises a bacteriophage coat and a DNA core that comprises plasmid DNA comprising: a) a host-specific bacteriophage packaging site wherein the packaging site is substantially in isolation from sequences naturally occurring adjacent thereto in the bacteriophage genome, b) a reporter gene, c) a bacteria-specific promoter operably linked to said reporter gene, d) a bacteria-specific origin of replication, and optionally e) an antibiotic resistance gene. The invention includes phage transducing particles, methods of making transducing particles, and methods of using the transducing particles in bacterial detection.

The invention provides a bacterium containing a polynucleotide comprising a nucleic acid encoding a heterologous antigen, as well as fusion protein partners. Also provided are vectors for mediating site-specific recombination and vectors comprising removable antibiotic resistance genes.

Provided herein are exemplary vectors for transforming algal cells. In exemplary embodiments, the vector comprises a Violaxanthin-chlorophyll a binding protein (Vcp) promoter driving expression of an antibiotic resistance gene in an algal cell. Embodiments of the invention may be used to introduce a gene (or genes) into the alga Nannochloropsis, such that the gene(s) are expressed and functional. This unprecedented ability to transform Nannochloropsis with high efficiency makes possible new developments in phycology, aquaculture and biofuels applications.

DNA-based methods employing amplification primers or probes for detecting, identifying, and quantifying in a test sample DNA from (i) any bacterium, (ii) the species Streptococcus agalactiae, Staphylococcus saprophyticus, Enterococcus faecium, Neisseria meningitidis, Listeria monocytogenes and Candida albicans, and (iii) any species of the genera Streptococcus, Staphylococcus, Enterococcus, Neisseria and Candida are disclosed. DNA-based methods employing amplification primers or probes for detecting, identifying, and quantifying in a test sample antibiotic resistance genes selected from the group consisting of blatem, blarob, blashv, blaoxa, blaZ, aadB, aacC1, aacC2, aacC3, aacA4, aac6'-lla, ermA, ermB, ermC, mecA, vanA, vanB, vanC, satA, aac(6')-aph(2''), aad(6'), vat, vga, msrA, sul and int are also disclosed. The above microbial species, genera and resistance genes are all clinically relevant and commonly encountered in a variety of clinical specimens. These DNA-based assays are rapid, accurate and can be used in clinical microbiology laboratories for routine diagnosis. These novel diagnostic tools should be useful to improve the speed and accuracy of diagnosis of microbial infections, thereby allowing more effective treatments. Diagnostic kits for (i) the universal detection and quantification of bacteria, and/or (ii) the detection, identification and quantification of the above-mentioned bacterial and fungal species and/or genera, and/or (iii) the detection, identification and quantification of the above-mentioned antibiotic resistance genes are also claimed.

The present invention relates to DNA-based methods for universal bacterial detection, for specific detection of the common bacterial pathogens Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Staphylococcus saprophyticus, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis as well as for specific detection of commonly encountered and clinically relevant bacterial antibiotic resistance genes directly from clinical specimens or, alternatively, from a bacterial colony. The above bacterial species can account for as much as 80% of bacterial pathogens isolated in routine microbiology laboratories. The core of this invention consists primarily of the DNA sequences from all species-specific genomic DNA fragments selected by hybridization from genomic libraries or, alternatively, selected from data banks as well as any oligonucleotide sequences derived from these sequences which can be used as probes or amplification primers for PCR or any other nucleic acid amplification methods. This invention also includes DNA sequences from the selected clinically relevant antibiotic resistance genes. With these methods, bacteria can be detected (universal primers and/or probes) and identified (species-specific primers and/or probes) directly from the clinical specimens or from an isolated bacterial colony. Bacteria are further evaluated for their putative susceptibility to antibiotics by resistance gene detection (antibiotic resistance gene specific primers and/or probes). Diagnostic kits for the detection of the presence, for the bacterial identification of the above-mentioned bacterial species and for the detection of antibiotic resistance genes are also claimed. These kits for the rapid (one hour or less) and accurate diagnosis of bacterial infections and antibiotic resistance will gradually replace conventional methods currently used in clinical microbiology laboratories for routine diagnosis. They should provide tools to clinicians to help prescribe promptly optimal treatments when necessary. Consequently, these tests should contribute to saving human lives, rationalizing treatment, reducing the development of antibiotic resistance and avoid unnecessary hospitalizations.

A Mob⁻ plasmid having a RSF1010 replicon, comprising a gene coding for Rep protein and said plasmid has been modified to inactivate gene related to mobilization ability. The present invention also describes a bacterium having an ability to produce useful metabolites, comprising the plasmid and said bacterium lack active thymidylate synthase coded by thyA gene and thymidine kinase coded by tdk gene, and a method for producing useful metabolites, such as native or recombinant proteins, enzymes, L-amino acids, nucleosides and nucleotides, vitamins, using the bacterium.

The invention provides a bacterial traceless genetic manipulation vector. The vector contains a toxin inverse-screening element controlled by an antitoxin switch and an antibiotics resistance gene. The invention also provides a construction method of the vector and a method for application of the vector in bacterial traceless gene knock-out, knock-in, replacement, point mutation and insertion and deletion of large DNA fragments (larger than 194kb). By the utilization of a constitutive promoter for continuous expression of toxin protein and by the utilization of an inducible promoter for inducible expression of antitoxin protein, the toxin inverse-screening element (TCCRAS) controlled by the antitoxin switch is established, and the problem that specificity of existing genetic manipulation systems of gram-positive bacterium is not high, the systems are instable and screening is difficult is solved.

The invention discloses a method for quickly reducing antibiotics and resistance genes in organic solid waste, and belongs to the field of organic solid waste treatment. The method has the advantagesthat ultrahigh-temperature aerobic fermentation is carried out on the organic solid waste by the aid of aerobic zymocyte capable of tolerating the temperatures of at least 80 DEG C under the conditionthat pile temperatures are not lower than 80 DEG C under the control for at least 5-7 days, and accordingly effects of quickly and stably reducing the antibiotics and the resistance genes in the organic solid waste can be realized; the antibiotics and the ARGs (antibiotic resistance genes) can be quickly degraded by means of ultrahigh-temperature aerobic fermentation, integral microbial communitystructures in piles further can be changed, 90% of microorganisms (mainly including Proteobacteria and Bacteroidetes) which carry the ARGs can be killed, risks of gene transfer of the ARGs can be reduced, diffusion of the ARGs can be controlled from the source, and the ARGs can be guaranteed against rebounding; the organic solid waste can be treated, and double effects of efficiently removing antibiotic residues and resistance gene pollution can be realized; exogenous heating can be omitted, energy can be produced only by the aid of metabolism of thermophilic microorganisms to reach the highfermentation temperatures, and accordingly the method is low in energy consumption and is environmentally friendly.

The present invention relates to DNA-based methods for universal bacterial detection, for specific detection of the common bacterial pathogens Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis, Streptococcus pneumoniae, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Staphylococcus saprophyticus, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis as well as for specific detection of commonly encountered and clinically relevant bacterial antibiotic resistance genes directly from clinical specimens or, alternatively, from a bacterial colony. The above bacterial species can account for as much as 80% of bacterial pathogens isolated in routine microbiology laboratories. The core of this invention consists primarily of the DNA sequences from all species-specific genomic DNA fragments selected by hybridization from genomic libraries or, alternatively, selected from data banks as well as any oligonucleotide sequences derived from these sequences which can be used as probes or amplification primers for PCR or any other nucleic acid amplification methods. This invention also includes DNA sequences from the selected clinically relevant antibiotic resistance genes. With these methods, bacteria can be detected (universal primers and/or probes) and identified (species-specific primers and/or probes) directly from the clinical specimens or from an isolated bacterial colony. Bacteria are further evaluated for their putative susceptibility to antibiotics by resistance gene detection (antibiotic resistance gene specific primers and/or probes). Diagnostic kits for the detection of the presence, for the bacterial identification of the above-mentioned bacterial species and for the detection of antibiotic resistance genes are also claimed. These kits for the rapid (one hour or less) and accurate diagnosis of bacterial infections and antibiotic resistance will gradually replace conventional methods currently used in clinical microbiology laboratories for routine diagnosis. They should provide tools to clinicians to help prescribe promptly optimal treatments when necessary. Consequently, these tests should contribute to saving human lives, rationalizing treatment, reducing the development of antibiotic resistance and avoid unnecessary hospitalizations.

The invention discloses an engineering bacterium through anthracene nucleus antibiotic and appliance, which is characterized by the following: inserting or replacing restriction enzymes site of dnmV gene nucleic acid order of sky blue rufus streptomycete with antibiotic against property gene and surface cerubidin synthetic gene; blocking-up engineering bacterium of dnmV gene; or transforming expressing carrier of anthracene nucleus antibiotic synthetic gene to lead streptomycin (S.lividans)TK24; extracting expressing plasmid from S.lividans TK24; leading-in sky blue rufus streptomycete gene only with antibiotic against property gene to block dnmV gene; blocking-up abrupt change bacterium. This invention can produce cerubidin and/or analogue, such as anthracene nucleus antibiotic.

The invention discloses an artificial wetland system capable of strongly removing an antibiotic and an antibiotic resistance gene from culture waste water, and relates to waste water treatment equipment which is an artificial wetland ecological treatment device capable of efficiently removing nitrogen and phosphorus elements, high-residual veterinary antibiotic contents and the antibiotics resistance gene from the livestock culture waste water. The system comprises a vertical subsurface flow artificial wetland device, adopts a filling manner of large size grading difference 'positive and negative particle size mixture' filling and paving, and takes red soil, waste bricks, and zeolite as main wetland fillings; the red soil, the waste bricks and the zeolite have large specific surface areas, good adsorptive capacity to nitrogen, phosphorus, the antibiotic and the antibiotic resistance gene, and high biological holding; and continuous, high-efficiency, and steady operation of the artificial wetland ecological system is ensured. The system has the advantages of low cost, low energy consumption, low possibility of blocking, high removal efficiency of the antibiotic and the antibiotic resistance gene, high efficiency of denitrification and dephosphorization, small occupied area, simple structure, convenient operation management and the like.

The invention provides a simple method for efficiently extracting DNA (deoxyribonucleic acid) from soil samples. According to the simple method, the DNA in environmental samples is fully released and exposed by using combination of methods such as glass bead grinding, liquid nitrogen repetitive freeze-thawing, SDS (sodium-dodecyl sulphate), lysozyme and a protease-K method for the different soil samples, the DNA purification is carried out through multiple phenol imitation extraction processes, and the determination of the DNA extraction efficiency is carried out by adding an internal standard bacterial strain Xcc8004. The simple method has simple requirements on sample pretreatment; the quantity of the required environmental samples is small; the obtained DNA has relatively high concentration and purity; the extraction efficiency is relatively high; the analysis and detection of antibiotic resistance gene pollution in the complex environmental samples can be met; and the method is simple and reliable and has practical application value.

The invention discloses a single-resistance escherichia coli-bacillus subtilis shuttle expression vector and application thereof. The bacillus subtilis shuttle expression vector provided by the invention comprises the following elements: a bacillus subtilis formed strong promoter P43, a sequence which encodes signal peptide, multiple cloning sites, an antibiotics resistance gene (kalamycin or chloramphenicol), a bifunctional promoter sequence which can start resistance gene expression of kalamycin or chloramphenicol in escherichia coli and bacillus subtilis, and copy homing sequences of bacillus subtilis and escherichia coli. The expression vector disclosed by the invention not only can be stably copied in escherichia coli, but also can be stably copied in bacillus subtilis, and expresses homologous and heterologous proteins. All vectors just contain one resistance gene, the plasmid of which is less than 400bp, so that gene operation and plasmid conversion are easy to carry out, and the conversion ratio is high. Besides the expression vector of an exogenous gene, the vector can be further used for screening promoters and constructing a gene knock-out vector.

The invention provides a bacterium containing a polynucleotide comprising a nucleic acid encoding a heterologous antigen, as well as fusion protein partners. Also provided are vectors for mediating site-specific recombination and vectors comprising removable antibiotic resistance genes.

The invention discloses a high-grade oxidization method for removing antibiotic resistance genes in sewage. In the provided method, the antibiotic resistance genes in sewage are removed by the following steps: filtering the sewage by grating, performing precipitation separation, carrying out Fenton oxidation, performing UV/H2O2 oxidation, disinfecting, and discharging. In the Fenton oxidation method, the influences of Fe2+/H2O2 mole ratio, H2O2 concentration, initial pH value and reaction time are all taken into account, and in the UV/H2O2 oxidation method, the influences of the illumination time, the addition amount of H2O2, pH value, and the like are considered so as to find the optimal operation parameters. The provided method can effectively remove the antibiotic resistance gene in sewage, and provides a novel technology for cutting off the propagation of antibiotic resistance genes in sewage.

The invention discloses a method for eliminating antibiotics resistance gene pollution in water by a hydrogen peroxide coupled ultrasonic method and belongs to the technical field of water treatment. The method provided by the invention is used for solving the problem that the water is polluted by the antibiotics resistance gene. The method is characterized in that the method of feeding hydrogen peroxide into water in combination with ultrasonic radiation is employed so that the antibiotics resistance gene is broken from the point of chemical reaction and basic groups, riboses and phosphoric acids in the antibiotics resistance gene are oxidized by means of strong oxidizing property of the hydrogen peroxide, and consequently, the chemical nature of the antibiotics resistance gene is changed and has no ability of storing and transferring genetic information, on one hand, and on the other hand, many cavitation bubbles are formed in water by means of ultrasonic. When the cavitation bubbles are broken, strong physical destruction effect is generated on the antibiotics resistance gene in water so that the double helix structure of the gene is decomposed and the nucleotide chain of the gen is broken; as a result, the genetic information contained in the nucleotide chain cannot be transferred; therefore, the purpose of eliminating the antibiotics resistance gene pollution is achieved.

In certain embodiments, the present invention concerns genotypic identification of bacteria that are resistant to a bacteria and subsequent determination of an appropriate therapy. In specific embodiments, a high-throughput genotypic detection method for biomarkers for antibiotic resistance and susceptibility allows efficient prescription practice and increases the likelihood of a successful therapeutic outcome. In certain embodiments, the information from the genotypic detection method is utilized for determining antibiotics that should be avoided or, alternatively, employed.

The invention discloses a method for removing antibiotic resistance genes through ionizing irradiation. Antibiotic mushroom dregs are treated with an ionizing irradiation technology (including gamma-rays and high-power electron beams generated by an electronic accelerator) to damage DNA of microbial cells, so that the resistance genes are effectively removed, and residual antibiotics can be decomposed simultaneously. The method is efficient and wide in application range; radiation can be performed at the normal temperature, no chemical reagents or only a few chemical reagents are required to be added, and no secondary pollution can be caused. The method can be applied to removal of the resistance genes in the antibiotic mushroom dregs, can be applied to removal of the resistance genes in water, soil and sludge and has wide application prospects in the environmental field.

The invention discloses a construction method of a gene engineering strain without plasmid and antibiotic resistance screening marker. A pXKF3T5b plasmid is used as the carrier to transform a target gene into a host cell, and then an auxiliary plasmid for expressing the integrase is removed, a kanamycin resistance gene in the integrated carrier is removed, the chromosome is induced by the triclosan to evolve and the evolved strain is fermented and screened, so as to obtain the gene engineering strain without plasmid but with high gene copy number; moreover, a screen marker of the pXKF3T5b plasmid is the fabI gene (for coding enoyl-acetyl carrier protein reductase) in Escherichia coli and the fabI gene is the triclosan resistance gene, thus the obtained gene engineering strain does not have the antibiotic resistance screening marker, furthermore can not cause the spread of the antibiotics drug-resistant bacteria in the environment and can be suitable for the industrialized production. In addition, the target gene of the gene engineering strain is integrated into the chromosome, thus the gene engineering strain can not be lost during sub cultures, the genetic stability is high, and the production performance is stable.

The present invention provides a method and components thereof of performing genetic modification under a drug-free environment. The method comprises the steps of generating a trapped mammalian cell library by trapper constructs (including the element of piggyBac terminal inverted repeats (TIRs)), reporter constructs, and helper constructs (including a sequence of an internal ribosomal entry site (IRES)). The present art allows: (1) to target & identify the silenced loci; (2) to separate genes with low-level expression at certain differentiation stages; (3) to evaluate the efficiency of gene targeting in the silent or repressed loci. The present invention avoids the biased gene targeting observed in the prior arts, and eliminates the needs of introducing antibiotic genes into the host genome which may lead to a potential threat of drifting antibiotic resistant genes into environment.