56 results about "Gyrb gene" patented technology

The invention relates to bacillus subtillis R31 with broad-spectrum antagonism capacity on plant pathogenic fungi, and in particular provides a bacterial strain (CCTCC NO: M209261) of the Bacillus subtillis R31 as well as growth inhibition action thereof on the plant pathogenic fungi and especially capacity thereof on the biological prevention and control of banana vasicular wilt. The plant endophytic bacillus subtillis is proved to be new bacillus subtillis through obtained 16S rDNA and gyrA and gyrB gene segments as well as BLAST search and comparison on NCBI (National Center of Biotechnology Information) and has better flat antagonism activity on multiple plant pathogenic fungi, better antagonism capacity especially on various soil-borne and vascular bundle diseases, i.e. fusarium oxysporum, rhizoctonia solani, alternaria, and the like, and better prevention and control effect in greenhouses and fields.

The invention relates to a bacillus subtilis str.TR21 (hereinafter referred to as 'TR21') strain having broad-spectrum antagonism to plant pathogenic fungi, in particular to the bacillus subtilis TR21 strain and a growth inhibition function thereof on the plant pathogenic fungi. The bacillus subtilis TR21 strain provided by the invention is preserved in China Center for Type Culture Collection (CCTCC) in January 20, 2010 with a preservation number of CCTCC No: M2010019, wherein the China Center for Type Culture Collection is located in Wuhan University, Lojia Mountain, Wuchang, Hubei, China. The strain provided by the invention is proved by obtained gene segments 16S rDNA, gyrA and gyrB and BLAST search and comparison on NCBI to be a novel bacillus subtilis strain having relatively higher flat antagonistic activity to a plurality of plant pathogenic fungi, particularly having high antagonism to various soil borne vascular diseases, such as fusarium oxysporum, rhizoctonia solani, alternaria and the like, and having good prevention and control effects shown in fields.

The present invention provides a gene chip for detecting common pathogen in dairy, which comprises a solid-phase vector and an oligonucleotide probe fixed on the solid-phase vector, wherein the oligonucleotide probe includes DNA fragment or its complementary DNA or RNA sequence selected from E. sakazakii, streptococcus pyogenes, staphylococcus aureus, Klebsiella pneumoniae, Klebsiella oxytoca, Listeria monocytogenes, salmonella and 16S-23S rDNA intergenic spacer region of Bacillus cereus, as well as ipaH gene of Shigella or gyrB gene of citrobacter freundii. The invention further provides a reagent box which uses the gene chip to detect pathogen in dairy. The gene chip and the reagent box of the invention for detecting pathogen in dairy are easy to operate, highly precise and greatly repeatable.

The invention discloses a gene chip of aquatic product cultivation pathogenic bacterium, comprising a solid phase carrier which is modified chemically, a detection probe and a quality control probe are distributed on the solid phase carrier in a dot matrix way; the detection probe comprises specificity 16S rDNA sequences and / or gyrB gene sequences of vibrio, comma bacillus, vibrio harveyi, vibrio alginolyicus, vibrio anguillarum, vibrio parahemolyticus, nocardia, nacardia seriolea, aeromonas, hydrophilic aeromonas, streptococcus and dolphin streptococcus, which are to be detected, the quality control probe includes PCR positive, chip fixed positive control, chip hybridizing negative control, chip hybridizing positive control and chip hybridizing blank control; the gene chip has the advantages of small volume and high flux, can detect known and unknown germs of the vibrio, the nocardia, the aeromonas and the streptococcus, and can detect specific germs with multiple kinds, and the simpleness and rapidness and specificity of the germs can be detected, and automatic detection can be carried out after detection software is additionally arranged.

The invention provides a gene chip of main pathogenic microorganism in drinking water and a testing kit, which mainly aims at 11 kinds of bacteria of colibacillus / Shigella, salmonella, vibrio cholera, vibrio parahaemolyticus, staphylococcus aureus, enterococcus faecails, pseudomonas aeruginosa, legionella pneumophilia, pneumobacillus, yersinia enterocolitica and the like, and L.interrogans. The gene chip comprises a solid phase carrier and a oligonucleotide probe fixed on the solid phase carrier, wherein the oligonucleotide probe contains gyrB gene with tremendous evolutionary advantage, ITS gene and DNA segment selected from 16srRNA gene or complementary DNA segment. The gene chip and the testing kit of the invention can test the main pathogenic microorganism in drinking water, and has the characteristics of simple operation, high throughput, high accuracy, strong repeatability and the like, and can be used for clinical test for the water quality monitoring department.

The invention relates to two ralstonia-solanacearum-resistant endophytic bacillus velezensis strains and application thereof. By separation, screening and purification in roots of healthy tobacco plants in a ralstonia solanacearum occurrence area of the Enshi Prefecture, two biocontrol bacillus endophyticus strains R-3 and R-9 are obtained, have evident resistance effects on ralstonia solanacearumand are both capable of generating high-temperature-resistant ralstonia-solanacearum-resistant substances. By gyrB gene sequence identification and phylogenetic tree construction, results show that the R-3 and R-9 are both bacillus velezensis; by potted plant root irrigation experimental verification, the bacilli R-3 and R-9 have great colonization performance in tobacco roots and can enter stemsthrough root transmission; by field experimental verification, bacillus velezensis R-3 and R-9 are effective in ralstonia solanacearum prevention and treatment, remarkably superior to chemical agentkasugamycin and free of pollution. The biocontrol endophyte research field is further enriched by researching on the bacilli R-3 and R-9, and a guiding significance to ralstonia solanacearum prevention and treatment is achieved.

The invention provides a gene chip for detecting pathogens of lower respiratory tract, which comprises a solid phase carrier and an oligonucleotide probe fixed on the solid phase carrier, wherein the oligonucleotide probe comprises a sequence selected from one or more DNA sequences of staphylococcus aureus, 16s-23s rDNA intergenic spacer region of Klebsiella pneumoniae or Acinetobacter baumannii, 16S gene of pseudomonas aeruginosa or haemophilus influenzae, gyrB gene of streptococcus pneumoniae or legionella pneumophila and copB gene of moraxella catarrhalis or the complementary DNA or RNA sequence. The invention further provides a reagent kit containing the gene chip. The utilization of the gene chip and the reagent kit can detect the pathogens of the lower respiratory tract; furthermore, the operation is simple, the accuracy is high and the repeatability is strong.

The invention provides a genetic chip and a detection kit related to major pathogenic microorganisms causing infectious diarrhea of human beings, which are mainly specific to 13 types of bacteria including lapactic bacillus coli / shigella, salmonella, comma bacillus, vibrio parahemolyticus, aeromona hydrophila, plesiomonas, virio hollisae, vibrio fluvialis, vibrio mimicus, Wauter's strains, vibriodamsela and vibrio furnissii. The genetic chip comprises a solid-phase carrier and an oligonucleotide probe fixed on the solid-phase carrier, wherein the oligonucleotide probe comprises DNA (Deoxyribonucleic Acid) fragments selected from genes including a gyrB gene, an ITS gene, a dnaJ gene, a toxR gene and the like with remarkable biological evolution advantages or complementary DNA fragments. By adopting the genetic chip and the detection kit provided by the invention, major pathogenic microorganisms causing infectious diarrhea of human beings can be detected. The genetic chip and the detection kit have the characteristics of easiness and convenience for operation, high flux, high accuracy, high repeatability and the like, and can be applied to clinical detection of inspection departments in hospitals.

The invention belongs to the fields of microbial technology and biological control and particularly relates to bacillus amyloliquefaciens BA-KA4 and a screening method and application thereof. Bacteria are separated from soil, and an agar disk diffusion method is adopted for screening BA-KA4 out of the bacteria, wherein the inhibition ratio of BA-KA4 on cucumber botrytis cinerea is 65%, and BA-KA4 has stable inheritance. Through cultural characteristics, morphology and physiological and biochemical characteristics of the strain in combination with result reanalysis of a 16S rDNA sequence and a gyrB gene sequence, it is preliminarily identified that the strain BA-KA4 is bacillus amyloliquefaciens. The strain is stored with the accession number of CGMCC No.12190 in the Common Microorganism Center of China General Microbiological Culture Collection Center. The strain has the best control effect on the cucumber botrytis cinerea, a long lasting period and a broad antibacterial spectrum.

The invention discloses a gene chip detecting marine pathogenic vibrios, and a preparation method and a detection method thereof. The gene chip comprises a solid-phase carrier and detection probes fixedly disposed on the solid-phase carrier, and the detection probes comprise vibrio vulnificus gyrB gene probe, vibrio vulnificus virulence gene hemA probe, vibrio splendidus gyrB gene probe, vibrio splendidus virulence gene toxR probe, vibrio harveyi gyrB gene probe, vibrio harveyi virulence gene hemA probe, vibrio parahaemolyticus gyrB gene probe, vibrio parahaemolyticus virulence gene toxS probe, vibrio anguillarum gyrB gene probe and vibrio anguillarum virulence gene hem probe which are respectively shown as SEQ NO 1-10. The gene chip is firstly applied to detection on marine pathogenic vibrios. The gene chip is capable of rapidly sensitively detecting target bacterium infectio, also is good in repeatability and strong in signal, and does not easily cause a nonspecific signal.

PCT No. PCT / JP97 / 00991 Sec. 371 Date Jan. 6, 1999 Sec. 102(e) Date Jan. 6, 1999 PCT Filed Mar. 25, 1997 PCT Pub. No. WO97 / 35970 PCT Pub. Date Oct. 2, 1997An oligonucleotide is provided which has a nucleotide sequence derived from SEQ ID NO:1, characterized in that it contains at least one site capable of amplifying a nucleotide sequence characteristic of Vibrio parahaemolyticus. The oligonucleotide may have a nucleotide sequence not derived from SEQ ID NO:3, or incapable of amplifying nucleotide sequences originating in Vibrio alginolyticus and Vibrio harveyi, and may be represented by SEQ ID NO:5 or SEQ ID NO:6. A method of detecting Vibrio parahaemolyticus in a specimen is also provided which comprises preparing a primer set comprising two of the above oligonucleotides, selectively amplifying therewith a DNA gyrase subunit B gene sequence contained in the specimen as a target, and determining whether or not there is a gyrB unit specific for Vibrio parahaemolyticus in the specimen. Also provided is a primer which reacts specifically with a gyrB gene of Vibrio parahaemolyticus to thereby differentiate and identify the same among other Vibrios and strains other than the genus Vibrio. The Vibrio parahaemolyticus-specific primer serves to detect 285-bp gyrB gene fragments specific for this Vibrio by the PCR method without the necessity for DNA extraction or like operations from bacterial cells.

The invention discloses a method and a detection kit for detecting a mycobacterium tuberculosis complex cluster based on a thermostatic technology. According to the method, 5 primers are designed aiming at a GyrB gene of mycobacterium tuberculosis, wherein TP is composed of an annealing sequence and a nucleotide sequence complementary with a target sequence; FP has a loop-stem structure, and can anneal with two complementary DNA single strands from two ends and stretch towards an opposite side; annealing sites of OP1 and OP2 are respectively located on the outer sides of the TP and the FP, anneal and stretch towards middle and simultaneously peel off double strands stretching from the TP and the FP; a single strand with the TP or FP, after being replaced, can serve as a template to continue the process, thus forming two key single-stranded intermediate products; the two intermediate products are then used for synthesizing DNA through self-guidance by taking special structures of the TP and the FP as starting points; through such circulation and repetition, a self-circulating chain displacement reaction is triggered under function of DNA polymerase, thus realizing massive amplification of the target sequence.

The invention relates to a primer for respectively amplifying specific regions of gyrB genes (DNAgyrase B subunit gene, hereinafter referred to as gyrB) in Legionella micdadei, Legionella bozemanii and Legionella longbeachae, but also provides a PCR (Polymerase Chain Reaction) reagent kit comprising the primer for amplifying the specific regions of gyrB genes in Legionella micdadei, Legionella bozemanii and Legionella longbeachae. The PCR reagent kit for detecting Legionella micdadei, Legionella bozemanii and Legionella longbeachae is simple, convenient and quick and has good specificity and high sensitivity, can be applied to the fields of supervision and detection of water bodies and clinic samples, detection of pathogenic bacteria in drinking water, bacteriology classification, epidemiology survey and the like and has profound and lasting social benefit and great economic benefit.

The invention relates to a primer for amplifying the gene specific area of a DNA gyrase B subunit gene (gyrB for short) in legionella pneumophilia and provides a polymerase chain reaction (PCR) kit which comprises the primer for amplifying the gene specific area of the gyrB in the legionella pneumophilia. The PCR kit is simple, convenient and rapid and has high specificity and sensitivity when used for detecting the legionella pneumophilia, can be applied to the fields such as the supervision and detection of water bodies and clinical samples, the detection of pathogenic bacteria in drinking water, bacteriology classification, the investigation of epidemiology and the like and has good social benefit and economic benefit.

The invention discloses a detection kit and detection method for flavobacterium columnare. The kit contains a pair of primers: an upstream primer GyrB-II-F and a downstream primer GyrB-II-R, designed by taking a GryB gene of the flavobacterium columnare as a target gene. The method can be used for fast and accurately detecting whether an alepidote body or a culturing water body contains pathogenic bacteria or not and achieves targeted fish disease prevention and control.

The invention belongs to the technical field of biology, and particularly relates to bacillus amyloliquefaciens and an application thereof. According to the invention, one strain T23 with strong antagonism to various pathogenic bacteria is separated and screened from various plant tissues and rhizosphere soil thereof by adopting dilution separation and plate confrontation culture methods. Throughmorphological and physiological and biochemical characteristic tests and 16s rRNA and gyrB gene sequence analysis, the bacterium is identified as bacillus amyloliquefaciens. The activity of metabolites of the strain is further determined, and the prevention and treatment effect of the antagonistic strain on tomato early blight, sclerotinia rot of colza and snakegourd fruit anthracnose is determined through a greenhouse pot experiment, so that the subsequent research and development of biocontrol agents for tomato early blight, sclerotinia rot of colza and snakegourd fruit anthracnose are convenient.

To produce high-performance specific gene amplification primers for detecting, quantifying, or identifying Vibrio cholerae and Vibrio mimicus, having low risk of misidentification and practically sufficient amplification efficiency and amplification specificity. We have determined partial nucleotide sequences of rpoD and gyrB genes of Vibrio cholerae, Vibrio mimicus, and closely related species, revealed their phylogenetic relationship, and then identified nucleotides characteristic of Vibrio cholerae and Vibrio mimicus, respectively. Thus, we have made it possible to design probes having high specificity and gene amplification primers having high specificity and excellent amplification efficiency, both of which contain the characteristic nucleotides.

The invention provides a kit and a detection method for rapidly detecting harveyi Vibrioharveyi. The kit and the detection method are designed, taking a pair of primers which are designed in harveyi Vibrioharveyi heat-resistant direct gyrB gene conservative area sequence as a main body. The invention adopts the polymerase chain reaction (PCR) technology and implements the qualitative detection on the specific DNA fragment of marine aquatic pathogen, the harveyi Vibrioharveyi. The invention has the advantages of convenient and rapid properties, good specificity and high sensitivity and can be applied to both the harveyi Vibrioharveyi tracking detection during the culture processes of various periods of aquatic animals and the environmental monitoring. With high practical value, the invention can avoid the transmission and the prevalence of germs.

The invention provides a detection kit for aeromonas veronii and an application method thereof. In the technical scheme, gyrB gene is found to be a single-copy house-keeping gene through experiments and has more remarkable advantages than conventional 16S rRNA in the distinguishing and identifying of aeromonas veronii and sibling species thereof; and meanwhile, Aha gene is found to be stably exist in the aeromonas veronii and related to the toxicity thereof. Based on the beneficial discoveries, the gyrB gene and Aha gene are adopted as target genes to develop a duplex PCR detection method for aeromonas veronii; in the method, specific primers are designed for the two target genes, and a PCR reaction system and reaction conditions are determined. In the invention, the sensitivity and specificity of aeromonas veronii detection are high, the pelteobagrus fulvidraco-source pathogenic aeromonas veronii can be quickly and accurately detected, and the wrong detection and missed detection are effectively avoided. Meanwhile, repeated culture and redundant series of biochemical reactions are prevented, and the time, labor and cost are saved.

The invention relates to a multiple PCR primer used for simultaneously detecting five pathogenic bacteria and three virulence genes in marine, and a designing method thereof. The method includes: designing a multiple PCR primer composition capable of simultaneously detecting enterococcus faecalis, edwardsiella tarda, carnobacterium, bacillus thuringiensis, vibrio alginolyticus, an enterobacter cloacae gyrB gene, a vibrio parahaemolyticus tlh gene, and an enterococcus faecalis cylA gene by utilization of a primer designing software, screening primers dominant in competitive states, subjecting the primers dominant in the competitive states to amplification and verification by utilization of a PCR method, and finally removing primers weak in the competitive states in the amplification primers so as to obtain a multiple PCR primer composition. A detection method of the multiple PCR is also disclosed. Compared with the prior art, the multiple PCR primer, the designing method and the detection method are advantageous in that: the multiple PCR primer composition simultaneously detecting the five pathogenic bacteria and the three virulence genes in marine has advantages of simple and convenient operation, rapidness, strong specificity, high sensitivity, and the like.

The invention discloses a loop-mediated isothermal amplification-based nucleic acid test method for Riemerella anatipestifer. The method includes the following steps: a specific primer sequence is designed according to the gyrB gene of the Riemerella anatipestifer, the DNA (deoxyribonucleic acid) of a sample to be tested is extracted as a template, and LAMP (loop-mediated isothermal amplification) reaction is carried out; an LAMP result is analyzed, and if the LAMP amplification result is positive, then the sample to be tested contains the Riemerella anatipestifer. The test method aimed at the Riemerella anatipestifer, which is provided by the invention, is simple, fast, specific, sensitive and economic, the result judgement method is simple, and the test method is suitable for use in clinic or grass-roots labs, and has a broad application prospect.

The invention discloses bacillus subtilis and application thereof to prevention and treatment of bipolaris maydis. The strain of the bacillus subtilis for preventing and treating bipolaris maydis is DZSY21, the preservation name is bacillus subtilis, the bacillus subtilis is preserved in the China General Microbiological Culture Collection Center (CGMCC) which is located in #3, No.1 Courtyard, West Beichen Road, Chaoyang District, Beijing, the preservation date is November 27, 2015, and the preservation No. is CGMCC NO.11749. The 16S rDNA gene sequence of the bacillus subtilis is a nucleotide sequence as shown in SEQ ID No.1, and the gyrB gene sequence of the bacillus subtilis is a nucleotide sequence as shown in SEQ ID No.2. The bacillus subtilis is high in growth speed, large in sporulation quantity, good in stress resistance and good in antibacterial property, and can be quantitatively colonized in leaves of corn plants.

The invention provides a multiplex PCR primer group and next-generation sequencing database building method for MLST (multilocus sequence typing) tracing of vibrio parahaemolyticus, and belongs to thetechnical field of microbiological detection. The multiplex PCR primer group for the MLST tracing of the vibrio parahaemolyticus comprises a primer for gyrB gene amplification, a primer for dnaE geneamplification, a primer for dtdS gene amplification, a primer for pyrC gene amplification, a primer for tnaA gene amplification, a primer for pntA gene amplification and one of a primer for recA-1 gene amplification and a primer for recA-2 gene amplification. The next-generation sequencing based database building method is established by multiplex PCR amplification for the seven genes, amplification and sequencing of the two types of RecA genes can be realized, and a sequencing result is relatively consistent with a first-generation sequencing result; and meanwhile, amplification of all genescan be realized by one-time PCR, the method can be directly applied to database building, and the operation is simple.

The invention discloses a general primer of cloned bacterium spiral-accelerating enzyme-gyrB gene, whose nucleic sequence of upstream primer is 5'-CC(ATGC)GG(ATGC)ATG TA(CT) AT(ATC)GG-3' and the nucleic sequence of downstream primer is 5'-CAT (CT) TC (ATGC) CC (ATGC) A (AG) (ATGC) CC (CT) TT (AG) (AT) A (ATGC) C (GT) (CT) TG-3'), wherein the whole sequence of PCR product approximates to the whole sequence of gyrB gene, which possesses stronger specificity and sensitivity.

The invention discloses a PCR (polymerase chain reaction) quick salmonella detection primer based on a gyrB gene, comprising the following gene sequences: S-P-for: 5'-GGTGGTTTCCGTAAAAGTA-3'; and S-P-rev: 5'-GAATCGCCTGGTTCTTGC-3'. The primer disclosed by the invention has the advantages of short detection period, reliable result, high sensitivity and the like, is simple to operate, is favorable for the early clinical diagnosis of all common salmonella diseases in people and animals, and has an important meaning for detecting the pathogenicity salmonella in public health, animal husbandry and veterinary and port quarantine.

The invention discloses a method for screening a bacterial strain for authenticating antagonistic aspergillus flavus. The method comprises the steps of aspergillus flavus spore suspension preparation,antagonistic bacterium preliminary screening, antagonistic bacterium secondary screening to obtain an antagonistic bacterium. The antagonistic bacterium is authenticated through morphological feature, physiological and biochemical feature, 16SrDNA and gyrB gene sequence analysis; the bacterial strain B10 with obvious inhibition on the aspergillus flavus is screened; through authentication, the bacterial strain B10 is bacillus amyloliquefaciens. A novel method and theoretical basis are provided for preventing and controlling aflatoxin B1 fungaltoxin poisoning. The method has important practical significance of production.

The phylogeny of twelve Campylobacter species was determined based on partial (1020-bp) gyrB gene sequences. Methods have been described for detection and speciation of Campylobacter, including 16S rRNA sequence analysis. However, gyrB provides a better resolution than the 16S rDNA gene for Campylobacter species with interspecies sequence similarities ranging from 58.3 to 89.2% compared to those reported for the 16S rRNA gene (ranging from 89 to 99%). A universal primer set, designed to amplify a 900-bp fragment of the gyrB gene in Campylobacter spp., was developed and used for PCR-RFLP of 19 strains representing twelve Campylobacter species and resulted in unique digest patterns for all twelve Campylobacter species. PCR assays for amplification of regions of the gyrB gene specific for each Campylobacter species were also developed. Using these PCR and PCR-RFLP methods results in unambiguous identification of the majority of Campylobacter species.

The invention relates to Bacillus subtilis strains for preparing fermented feather meal, and an application of Bacillus subtilis strains. An object of the present invention is to improve the hydrolyzed feather powder by adding Bacillus subtilis to the hydrolyzed feather powder for 1 days and increasing the digestibility of the feather powder with pepsin. Another object is to provide a Bacillus subtilis strain comprising a protease and a keratinase, which can be used in animal feed additives to enhance nutritional value. Strains of Bacillus subtilis 531 to 7 were isolated from the compost soiland compared by the degradability and enzyme activity test of feathers to confirm commercial value. Strains of Bacillus subtilis 531-7 were aligned by 16S rDNA sequencing and gyrB gene sequencing to confirm their novelty.