778 results about "Enterobacteriales" patented technology

A DNA encoding 20K hGH is connected directly to a gene encoding Escherichia coli OppA protein secretion signal, or a modified form thereof, and a DNA encoding signal peptidase 1 to construct a recombinant plasmid, E. coli is transformed by the plasmid and cells of the resulting E. coli transformant strain are cultured for secretory production of the 20K hGH in the E. coli periplasm. This method enables efficient secretory production of 20K hGH and easy isolation and purification of 20K hGH from the periplasm fraction because the level of impure proteins in the E. coli periplasm is low.

The inventive method allows peptides or polypeptides to be exposed on the surface of gram-negative host bacteria using specific intimin-based anchor modules. Intimins with shortened carboxy terminals have been found to be particularly suitable anchor modules for passenger domains in the exterior E. coli cell membrane. According to the method, host bacteria are transformed using vectors, on which are located a fused nucleic acid sequence consisting of a sequence segment which codes for an intimin with a shortened carboxy terminal and a nucleic acid sequence segment which codes for the passenger peptide that is to be exposed. The invention permits a particularly large number of passenger domains to be exposed on the cell surface of the bacteria, without adversely affecting the viability of the bacteria.

The invention relates to the field of microbial detection and discloses a method for rapid detection of enterobacter sakazakii. The method comprises the steps of: adding immunized superparamagnetism nanometer magnetic beads into a sample, adding a stabilizer, and incubating for 45-60min at 30-40 DEG C, wherein the magnetic beads are specifically bonded and enriched on microbial food-borne pathogenic bacteria, namely the enterobacter sakazakii, in the sample; and measuring spinning-spinning relaxation time and calculating a value deltaT2 by taking a bacteria-free sample as a blank control. The method disclosed by the invention has high specificity and sensitivity to the detection of the enterobacter sakazakii, is rapid, and is particularly suitable to detection of low-concentration enterobacter sakazakii, especially the enterobacter sakazakii in dairy products; in addition, the method is convenient and simple to operate and has good reliability and low requirement on assorted equipment.

Methods and compositions are provided for treating weight related conditions and metabolic disorders by altering microbiota in a subject. One aspect provides methods and compositions to alter microbiota in a subject by administering to the subject a composition that includes a substantially purified microbiota from phyla such as Bacteroidetes, Proteobacteria, Firmicutes and Verrucomicrobia or orders such as Bacteroidales, Verrucomicrobiales, Clostridiales and Enterobacteriales or genera such as Alistipes, Clostridium, Escherichia, and Akkermansia. Another aspect includes a pharmaceutical composition for altering microbiota that includes a therapeutically effective amount of substantially purified microbiota and a pharmaceutically acceptable carrier. Yet another aspect includes methods for treating a disorder, such as obesity, in a subject in need of such treatment by changing relative abundance of microbiota in a gastrointestinal tract of the subject without or in addition to a surgical procedure.

The invention discloses a kit for quickly detecting 15 pneumonia pathogenic bacteria. The kit can detect streptococcus pneumoniae, staphylococcus aureus, haemophilus influenzae, mycoplasma pneumoniae, pseudomonas aeruginosa, baumanii, enterococcus faecalis, enterococcus faecium, klebsiella pneumoniae, escherichia coli, enterobacter cloacae, stenotrophomonas maltophilia, burkholderia cepacia, legionella pneumophila and chlamydia pneumoniae which cover clinically common pneumonia pathogenic bacteria difficult to culture. 16S rDNA and specific gene sequences corresponding to the pneumonia pathogenic bacteria are detected by combining gene chips with multiple asymmetric PCR reactions, and the categories of the bacteria in a to-be-detected sample are identified in genus and species. The kit makes up for the defect that current clinical detection of pneumonia pathogenic bacteria is not in time or comprehensive and a novel detection means for early diagnosis and early treatment of patients suffering from pneumonia is provided.

The invention discloses recombinant escherichia coli using glucose for producing hydroxytyrosol as well as a building method and application. The method comprises the following steps that SEQ ID NO.1 and 2 are used as primers; the escherichia coli BW25113 genome DNA (deoxyribonucleic acid) is used as a template; HpaBC gene PCR (polymerase chain reaction) augmentation is carried out; vectors pTrcHisB are connected; middle plasmids are obtained; the middle plasmids are used as templates; the SEQ ID NO.3 and 2 are used as primers; PCR augmentation is carried out, and plasmids pTrcHisB-ARO10 are connected; expression vectors pTrcHisB-ARO10-HpaBC are obtained; in addition, the vectors and the plasmids pBb3 are simultaneously transferred into escherichia coli strains BMGA to obtain the recombinant escherichia coli. The recombinant escherichia coli contains ARO10 genes from saccharomyces cerevisiae, and HpaBC genes from the escherichia coli endogenesis. The yield of the hydroxytyrosol can be obviously improved.

Disclosed herein are various open reading frames from a strain of E. coli responsible for neonatal meningitis (MNEC), and a subset of these that is of particular interest for preparing compositions for immunising against MNEC infections.

The invention discloses a method for simultaneously and quickly detecting salmonella typhimurium, escherichia coli O157:H7 and listeria monocytogenesis in food. The method comprises the following steps of: preparing nano immunomagnetic particles of the targeted typhimurium, escherichia coli O157:H7 and listeria monocytogenesis so as to capture target bacteria from a detection sample; obtaining the target bacteria by magnetic field separation; treating by adopting propylmercuric bromide azide (PMA) to eliminate the interference of killed bacteria; and finally, extracting DNA (Deoxyribose Nucleic Acid) to carry out multiple PCR (Polymerase Chain Reaction) detection. Compared with a classical bacteria separation and identifying and serological typing method, the method disclosed by the invention is quick and accurate, can be used for only detecting viable bacteria and can be used for simultaneously detecting and identifying the three pathogens.

The bacterial colony number sample for verifying food microbiological capacity relates to quality control technology in microbiological detection. The sample is matrix with added Bacillus cereus as leading bacteria and background bacteria. The matrix is practical food, and the background bacteria include one or several kinds of serratia marcescens, Citrobacter freundii, enteroaerogen and escherichia coli. The sample of the present invention has homogeneous colony number content, high finishing rate, and high stability, and may be used in test capacity verification of domestic and international labs. In the same time, the capacity verifying sample may be used in verification of detection method, calibration of test instrument, quality control and check of test result and other aspects.

This invention relates to a system for the expression of cytochrome P-450 gene in host Escherichia coli, and provides Escherichia coli which contains actinomycete ferredoxin gene and also ferredoxin gene and ferredoxin reductase gene which are xenogenic to Escherichia coli. Thus, this invention is useful for the promotion of effective single oxygen atom insertional reaction of a substrate organic compound.

An Escherichia coli recombinant strain producing shikimic acid, and a construction method and application thereof belong to the technical field of microbial gene engineering. The invention firstly utilizes a molecular biology technique to delete shikimic acid kinase I gene (aroK) and shikimic acid kinase II gene (aroL) of Escherichia coli CICIMB0013, and a gene (ptsG) of a key protein EIIBC<Glc> and a quinin acid / shikimic acid dehydrogenase gene (ydiB) of a glucose phosphotransferase system to obtain an Escherichia coli mutant strain CICIMB0013.SA4 (delta aroK, delta aroL, delta ptsG, delta ydiB). The invention also constructs a recombinant expression plasmid pTHGAA containing key genes comprising aroG*,ppsA and tktA in a metabolic pathway of shikimic acid; and the recombinant expression plasmid pTHGAA is transferred into the recombinant strain CICIMB0013.SA4 to obtain a recombinant Escherichia coli B0013 (SA4 / pTHGAA) capable of producing shikimic acid efficiently. The Escherichia coli recombinant strain provided by the invention can realize efficient accumulation of shikimic acid in a fermentation process.

The invention belongs to the technical field of biology detection and relates to an escherichia coli O1, O2, O18 and O78 serotype detection kit and a detection method thereof. Forward primers of a primer group of the kit are highly conservative escherichia coli O antigen combined relative gene sequences, and backward primers are four serotype specific primers designed on the basis of four O1, O2, O18 and O78 serotype escherichia coli O antigen synthesis relative genes. The detection kit containing the primer group and the detection method of the detection kit have the advantages of being fast, sensitive, specific, low in cost, easy to operate and capable of well overcoming the shortcomings in the current escherichia coli O1, O2, O18 and O78 serotype detection method, can meet the requirements for the current escherichia coli O1, O2, O18 and O78 serotype detection, can be popularized and used easily in a wide range and have wide market prospect and large economic benefit.

The invention discloses a preparation method and a drug carrying method of an escherichia coli outer membrane vesicle, and application of the outer membrane vesicle in anti-tumor. The preparation method of the escherichia coli outer membrane vesicle comprises the following steps of culturing a culture medium using LB in vitro, culturing an escherichia coli BL21 strain, performing centrifugation, filtration and ultrafiltration treatment to obtain an upper layer culture solution free of escherichia coli, and finally centrifuging the upper layer culture solution by a supercentrifuge to prepare the escherichia coli outer membrane vesicle. The particle size of the prepared escherichia coli outer membrane vesicle is uniform, and is about 50nm. Usually, a bacterium outer membrane vesicle is low in yield and difficult in drug carrying. The invention excogitates a novel drug carrying method for the bacterium outer membrane vesicle, so that the carrying efficiency of an anti-tumor drug can be significantly improved; an inhibition effect on multiplication and invasion of a tumor cell is increased; and a good application prospect of the bacterium outer membrane vesicle serving as a novel non-virus drug carrier is presented.

The invention relates to a method for detecting bacteria in drinking water and surface water, especially a method for simultaneous specific detection of bacteria from the Legionella species and the Legionella pneumophila species by in situ hybridization. The invention also relates to a method for specific detection of faecal streptococci by in situ-hybridization and a method for simultaneous specific detection of coliform bacteria and bacteria of the Escherichia coli species, in addition to corresponding oligonucleotide probes and kits enabling said inventive method to be carried out.

The invention discloses an enterobacter cancerogenus for degrading polyethylene and an application of the enterobacter cancerogenus. The bacterial strain disclosed by the invention is enterobacter cancerogenus YZ1 CGMCC No.6816. The active component of a microbial agent disclosed by the invention is enterobacter cancerogenus YZ1 CGMCC No.6816. The bacterial strain disclosed by the invention is capable of effectively corroding and degrading polyethylene plastic. The microbial agent disclosed by the invention is low in production cost, and can degrade 5% (mass percent) of polyethylene within 60 days. The bacterial strain and the microbial gent disclosed by the invention can be suitable for polyethylene wastes in refuse landfills and various water body and soil environments, and can ensure that the polyethylene can be biodegraded. According to the method disclosed by the invention for efficiently degrading microorganisms of plastic, the microorganisms of degradable plastic with high purity and high activity can be quickly obtained. A technique and a method can be provided for solving the problems that the existing polyethylene plastic wastes pollute the environment and occupy great landfill capacity of the landfill, and the enterobacter cancerogenus is of great importance in environmental protection and reduction of garbage disposal cost.

The invention discloses enterobacter cloacae and application of the enterobacter cloacae. The enterobacter cloacae SXH-1 provided by the invention has the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.6296. The invention also protects the application of the enterobacter cloacae SXH-1 to the plant growth promotion. The plant is concretely chives. The enterobacter cloacae SXH-1 provided by the invention can use ACC (aminocyclopropane carboxylic acid) as the unique nitrogen source for growth, and meanwhile, the ACC is decomposed. The strain is hopeful to realize important effects in the chive growth promotion aspect.

The invention discloses a non-induced expression genetic engineering strain and the constructing method and the application. The host cell of the strain is enteric bacilli E.coliBL21, CCTCC M205136, PCR expanding hrp Z gene section to clone to pGEM-T carrier, ferment cutting and bonding by EcoR I / Xho I, inserting the hrp Z gene into the enteric bacilli expression carrier pET-32b(+) lower reaches of thioredoxin, constructing rebuilding expression particle pET-32b-hrp, and transferring into E.coliBL21, filtering to gain positive clone. The non-induced expression rebuilt albumen HrpZ, SDS-PAGE shows that high efficiently expresses APDZ albumen. The rebuilt albumen has good prevention effect to plant disease and could improves the yield.

The invention discloses a preparation method of a nano-silver composite antibacterial agent. The invention is characterized in that the nano-silver composite antibacterial agent is formed by combining a nano-silver solution and an organic antibacterial agent which have synergistic effect and adduction effect according to a certain ratio. The structure and performance of the antibacterial agent after combination are stable, and the antibacterial agent has more rapid, efficient and broad-spectrum antibacterial performance. The minimal inhibitory concentration (MIC) of the antibacterial agent for inhibiting escherichia coli, Enterobacter hormaechei, staphylococcus aureus, bacillus subtilis, Rhodotorula mucilaginosa, penicillium and Aspergillus niger is only 1-10 microgram / ml. The preparation method of the nano-silver composite antibacterial agent is characterized in that the nano-silver in the nano-silver composite antibacterial agent has monodispersity and there is no agglomeration phenomenon. The preparation method has advantages of ingenious conception, simple experimental operation, high operationality and short period, and is suitable for large-scale production.

Compositions and methods for stimulating an immune response against a secreted enterohemorragic Escherichia coli (EHEC) antigen are disclosed. The compositions comprise EHEC cell culture supernatants.

The present invention discloses a construction method of escherichia coli for synthesizing fucosylated oligosaccharide by fermentation. The construction method includes the steps: (1) overexpressing at least one gene for encoding enzymes required for denovo synthesizing GDP-L-fucose in a prokaryotic host cell; (2) expressing an exogenous gene for encoding a fucosyltransferase in the prokaryotic host cell; (3) reducing or eliminating the activity of GDP-mannanohydrolase in the prokaryotic host cell; and (4) reducing or eliminating the activity of beta-galactosidase in the prokaryotic host cell.According to the present invention, the escherichia coli is constructed by using the method, and the escherichia coli may be used for preparing the fucosylated oligosaccharide. Shown by a result generated by performing fermentation production verification on the fucosylated oligosaccharide (with 2'-fucosyllactose as an example) in a 5L tank by adopting an engineering strain constructed by the present invention, the highest production level of the 2'-fucosyllactose (2'-FL) may reach 50 g / L.

The invention relates to a bacterium for the production of genetically-engineered subunit oil-adjuvant vaccines against infectious bursal disease, i.e. Escherichia coli BL21 / pET-28a-VP2, CCTCC No: M204038. The process for constructing the bacterium consists of inserting VP2 protein gene of the infectious bursal disease virus onto expression plasmid pET-28a to construct prokaryotic expression plasmid, then using the prokaryotic expression plasmid to transform the escherichia coli BL21, using lactose as inducer to induce Escherichia coli BL21 / pET-28a-VP2 expression VP2 protein, purifying and charging oil-adjuvant to obtain the vaccines.

This invention relates generally to a method for purifying Interleukin-4 or related variants from an Escherichia coli culture medium, where Interleukin-4 and its derivatives are expressed as insoluble protein aggregates, so-called inclusion bodies.

The invention belongs to the technical field of escherichia coli phage, and particularly relates to a broad-spectrum escherichia coli phage capable of cracking four bacteria and an application of the broad-spectrum escherichia coli phage to sterilization and bacterium prevention. The invention mainly discloses an escherichia coli phage EC35P1 (Escherichia coli phase EC35P1), and the preservation number of the escherichia coli phage EC35P1 is CCTCC M 2020438. The phage is a virulent phage separated from the nature, tests prove that the phage has no toxic effect on normal microbial flora, and DNA of the phage cannot encode virulence genes and is high in stability. The phage is wide in host range, can be used for cracking escherichia coli, shigella, salmonella and enterobacter cloacae, and large-scale industrial production can be realized. The phage provides an excellent strain resource for developing a novel antibacterial preparation, and has a good application and development prospect.

A method of preparing high-performance M-MLV reverse transcriptase is disclosed. The method includes a step of performing mutant cloning construction, with plasmid sequences of constructed mutants being sequenced for confirmation to obtain seventeen mutant plasmids; a step of mutant screening, namely a step of transforming wild-type pET28b-M-MLV and the seventeen mutant plasmids into escherichia coli competent cells, culturing, inducing mutant enzyme expression, collecting bacterium cells to obtain a mutant enzyme coarse extract liquid, and subjecting mutant enzyme coarse extract liquid having protein expression to reverse transcription activity screening at different temperatures; and a step of culturing the mutants positive in activity screening in a liquid medium, inducing protein expression, collecting bacterium cells, and performing purification to obtain the M-MLV reverse transcriptase. A combination of molecular rational design and function screening is adopted by the method, the M-MLV reverse transcriptase having high thermal stability can be obtained only by a small range of screening, and the thermal stability is that the M-MLV reverse transcriptase can resist a temperature of 65 DEG C.

The present invention relates to a novel bacteriophage having an E.-coli-specific bactericidal activity, a composition for the prevention or treatment of infectious diseases caused by Enterotoxigenic E.-coli comprising the bacteriophage as an active ingredient, an antibiotic comprising the bacteriophage as an active ingredient, a feed additive composition comprising the bacteriophage as an active ingredient, a sanitizer or cleaner comprising the bacteriophage as an active ingredient, and a method for treating colibacillosis using the bacteriophage. The novel bacteriophage of the present invention has a specific bactericidal activity against pathogenic E. coli, and excellent acid- and heat-resistance. Therefore, the novel bacteriophage can be used for the prevention or treatment of swine colibacillosis, which is an infectious disease caused by pathogenic E.-coli, and can also be widely used in animal feed additive compositions, sanitizers, and cleaners.

The present invention relates to a microbial cell sensor for detecting arsenic bioavailability degree, which is suitable for detecting bioavailability degree of arsenic in water and soil. The microbial cell sensor comprises: Escherichia coli as a host cell carrying recombinant plasmid. The recombinant plasmid is pUC18 plasmid containing arsenic resistant system ars promoter, arsenic resistant system controlling gene arsR, luciferase gene luc and rrnb terminator tandem sequence. The response time of the sensor on the arsenic is 30 minutes, the lowest detection concentration of the arsenic is 0.01 micromole / L, the highest detection concentration is 100 micromole / L. The microbial cell sensor has features of high sensitivity, fast response speed, low cost, and simple operation, and can be widely used in detecting the arsenic polluting the environment and evaluating the risk.

The invention discloses an Enterobacter mori and a method for producing natural vanillin by biotransformation of ferulic acid by the Enterobacter mori, which belongs to the microbial fermentation technical field. The method for producing natural vanillin by biotransformation of ferulic acid by Enterobacter mori CCTCC No: M2012020 comprises the following steps: 1)activating bacterial classification; 2) preparing a seed nutrient solution; 3) fermenting; 4) conversing, performing centrifugal treatment on a broth for 12 minutes under condition of 7000rpm / min, and collecting a supernatant to obtain the natural vanillin broth having light yellow color. The Enterobacter mori CCTCC No: M2012020 which is not reported has the advantages of low cost of carbon source and nitrogen source, and wide source, in the broth, the natural vanillin has the advantages of large concentration and high output, the production technology has the advantages of simple process, mild condition, friend environment and industrial prospect, and has good social benefit and economic benefit for changing waste into valuables.

The invention discloses a cutinase producing gene engineering bacteria and use thereof and belongs to the technical field of bioengineering. Recombinant plasmid Tfu-0883-hlyAs / pET20b(+) is constructed, Escherichia coli E.coliBL21(DE3) is converted to obtain recombinant Escherichia coli Tfu-0883-hlyAs / pET20b(+) / E.coliBL21(DE3). A certain specific growth speed is controlled in a feed supplement batch fermentation manner, fermentation culture is performed for 30 to 34 hours, and the enzyme survival rate in fermentation supernate reaches 700 to 750U / Ml. In the invention, glycerol is used as a main raw material, a semisynthetic culture medium is adopted, and the cutinase producing gene engineering bacteria have the advantages of stability, convenient regulation and the like and are suitable for large-scale production.

The invention relates to a genetic engineered bacterium which contains a recombinant plasmid pET28a-GMAS, is 6585 bp in a plasmid size, is 1308 bp in a fragment size of a GMAS, is named as Escherichia coli and has a kanapenecilin resistance. Genes of the [gamma]-glutamylmethylamine synthetase are inducibly expressed by isopropylsulpho-[beta]-D-galactoside. The genetic engineered bacterium has a capability of biologically synthesizing theanine. By means of glutamic acid and ethylamine as substrates, a method in the invention is simple in operation method, is convenient to control in production, is high in conversion and production efficiency of the theanine and has an excellent industrial application prospect.

The invention belongs to the technical field of biology and particularly relates to a riemerella anatipestifer-escherichia coli shuttle expression vector as well as a construction method and an application thereof. The riemerella anatipestifer-escherichia coli shuttle expression vector disclosed by the invention has a nucleotide sequence of SEQ ID NO:1. The invention further discloses a construction method and the application of the riemerella anatipestifer-escherichia coli shuttle expression vector. The riemerella anatipestifer-escherichia coli shuttle expression vector pRES constructed by the construction method can stably exist and can be copied in riemerella anatipestifer and escherichia coli; an exogenous gene carried by the riemerella anatipestifer-escherichia coli shuttle expression vector can be stably expressed in the riemerella anatipestifer; the riemerella anatipestifer-escherichia coli shuttle expression vector can be used for complementary assay of riemerella anatipestifer deletion mutation strains, research of recombinant riemerella anatipestifer vaccines and the like and has a wide application prospect.