728 results about "Enterobacteriales" patented technology

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.

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.

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.

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 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 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 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.

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 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 respiratory syncytial virus vaccine, and the preparation method and application, in particular to an application of escherichia coli to express and recombinant respiratory syncytial virus G protein and mutation G protein, and preparation vaccine with nontoxic typed escherichia coli heat labile enterotoxin, for human or animal preventive inoculation, and for respiratory syncytial virus resistance, belonging to the field of biotechnology. The respiratory syncytial virus vaccine is characterized in that: the respiratory syncytial virus subunits vaccine comprises lopped respiratory syncytial virus RSV protein G, and further comprises nontoxic typed escherichia coli heat labile enterotoxin LT adjuvant. The vaccines are lopped respiratory syncytial virus RSV protein G containing amino acid between aa130 and 230 of the original G protein, and substitutes the amino acid CAWIC (CX3C module ordered) between aa182 and 186 with the amino acid YLEKESIYY (CTL epitope) on the RSV M protein, forming the GCIL protein. The respiratory syncytial virus vaccine has the advantages of remarkable practical significance for preventing human respiratory syncytial virus infection.

The invention relates to a Qbeta-2aa phage virus-like particle protein which is prepared according to a method comprising the following steps of: (1) mutating a Qbeta phage CP (coat protein) gene terminator codon into a strong terminator codon TAA from TGA and then cloning the strong terminator codon into a protokaryon expression vector pET28a(+) to obtain a coding CP plasmid pETQB-CP; (2) inserting the nucleotides AAGCTT of coding lysine and leucine at the coding immunodominance determining region position in a Qbeta phage CP extension protein gene, i.e. between the 72th codon and the 73th codon of the Qbeta phage CP extension protein gene, and mutating the CP gene terminator codon into GGA from TGA and then cloning the GGA into a protokaryon expression vector PGEX4T-2 to obtain a codingA1 protein plasmid pGEXQbeta-A1; (3) jointly converting the mutated coding Qbeta phage CP plasmid and the A1 protein plasmid into escherichia coli, and inducing and then expressing to obtain the virus-like particle protein Qbeta-2aa; and (4) coupling a short peptide antigen to Qbeta-2aa phage virus-like particles by using a iso-bifunctional cross-linking agent.

The invention discloses a bacteria strain FY4-2 selected from mass gen-seng endophytic bacterias; the bacteria strain FY4-2 produces auxin, azotification, phosphate dissolving, potassium releasing and ironophore, and has growth promotion function to the host gen-seng, and is identified to be enterobacterasburiae; the bacteria strain FY4-2 is strong in IAA production capacity, can reach 13.5 ug/mL, has good growth vigor under nitrogen-free condition, has azotification potential, phosphate dissolving content can be measured in quantity to be 130.6ug/mL, potassium releasing can obtain 4.33ug/mL, and can produce ironophore with activity at 92%; the bacteria strain FY4-2 can promote gen-seng seed to germinate and grow, bud length can be increased by 81.97%; the bacteria strain FY4-2 has obvious promotion effect for gen-seng root growth, fresh weight can be increased by 40.5%, and dry weight can be increased by 43.97%; the bacteria strain FY4-2 is found to provide excellent basis for developing and using gen-seng growth promotion bacterial manure.