30 results about "Pseudomonas plecoglossicida" patented technology

The invention discloses a method to produce ADI strain by screening and the application of the method, pertaining to the field of bioengineering technology. The invention particularly relates to a rapid method to screen ADI strain and produce Pseudomonas plecoglossicida (CGMCC No.2039) of ADI, as well as a method to produce ADI by using the microorganism. Under an aerobic condition and an environment maintained at pH6.5-8.0, enzyme activity can achieve 1.6U / mL fermentation broth after 20-24h fermentation culture. ADI produced with such microorganism has very strong inhabitation agaisnt liver cancer cell strain (HepG2). ADI genes can be obtained from the chromosome DNA of such microorganism, and the gene order can be achieved.

The invention discloses a fluorescent quantitative PCR detection kit of Pseudomonas plecoglossicida, and a detection method thereof. The kit comprises 10.0mul of an SYBR Premix Ex TaqTMII reagent, 0.8mul of an upstream primer solution having a concentration f 10muM, and 0.8mul of a downstream primer solution having a concentration of 1.0muM. The detection method comprises the steps of bacterium extraction, bacterium enzymatic hydrolysis, DNA crude extract preparation, DNA crude extract purification and detection. The detection kit can sensitively, rapidly, quantitatively and specifically detect early-stage Pseudomonas plecoglossicida, and the qualitative and quantitative sensitivities can reach 10<3>copies; and the detection kit can detect whether aquatic farming animals are infected with the Pseudomonas plecoglossicida nor not, the content of the Pseudomonas plecoglossicida in the farming water environment, whether a farming feed and a fresh bait are polluted by the Pseudomonas plecoglossicida or not, and the like.

The invention provides a Pseudomonas plecoglossicida fliA gene silenced strain that is Pseudomonas plecoglossicida fliA-RNAi and deposited in the China Center for Type Culture Collection on November 12th, 2018 with an accession number of CCTCC NO:M 2018770. The constructed Pseudomonas plecoglossicida fliA gene silenced strain has virulence significantly lower than that of a Pseudomonas plecoglossicida wild strain, time of death caused by same infection conditions is delayed, and the death rate is greatly reduced, thus laying a theoretical foundation for development of an attenuated Pseudomonasplecoglossicida vaccine.

The present disclosure relates to a pseudomonas mutans and application thereof. The deposit number of Pseudomonas plecoglossicida of the present disclosure is CGMCC No.14475. The present disclosure also provides a microbial bacterial agent, which contains the above-mentioned Pseudomonas mutans as an active ingredient. The present disclosure also provides the application of the above-mentioned Pseudomonas mutans and the above-mentioned microbial bacterial agent in degrading sulfamethazine, treating poultry and livestock breeding waste and remediating antibiotic pollution. Through the above technical solution, the present disclosure can remove sulfamethazine more effectively.

The invention discloses a Pseudomonas plecoglossicida and an application thereof, and specifically provides a Pseudomonas plecoglossicida BD, whose preservation number is CGMCC No.14205. The embodiment of the present invention provides Pseudomonas plecoglossicida (Pseudomonas plecoglossicida) BD, which can grow and reproduce with nitrile compounds as the only energy source, can degrade pollutants such as nitrile compounds in acrylonitrile wastewater, and reduce the chemical oxygen demand of acrylonitrile wastewater Amount and biochemical oxygen demand, thereby improving the biodegradability of acrylonitrile wastewater.

The invention discloses a pseudomonas plecoglossicida DNA vaccine and a preparing method and application thereof. The pseudomonas plecoglossicida DNA vaccine is characterized in that the pseudomonas plecoglossicida DNA vaccine is pCI-pcrV; pseudomonas plecoglossicida NB2011 genome DNA serves as a template, and is subjected to PCR amplification through a primer P1 / P2, the amplified products are purified to be connected with plasmid pCI-neo, and recombinant plasmid pCI-pcrV is obtained, and is the pseudomonas plecoglossicida DNA vaccine, wherein the sequence of a target gene pcrV is shown in SEQID NO:1. According to the pseudomonas plecoglossicida DNA vaccine, the immune protective rate of pseudomonas plecoglossicida infection is 70% or above, and therefore the high protective rate is achieved. The immune protective rate of the pseudomonas plecoglossicida DNA vaccine in 8 weeks after immunization is not obviously reduced, and therefore the pseudomonas plecoglossicida DNA vaccine has thelong-acting advantage.

The present invention relates to a phenol-degrading bacterium producing biological surfactant and its application. Its technical scheme is: the provided bacterial strain is Pseudomonas plecoglossicida BPH-3, which is preserved by the China Center for Type Culture Collection, and the preservation number is: CCTCC NO: M 2019144, and the preservation date is March 14, 2019 , the GenBank accession number of the 16S rDNA of the Pseudomonas plecoglossicida (Pseudomonas plecoglossicida) BPH-3 is MK575471, and the Pseudomonas (Pseudomonas plecoglossicida) BPH-3 is a gram-negative bacteria, and the colony is milky white, circular , smooth surface, neat edges; negative under the microscope after Gram staining, the strain was short rod-shaped. The phenol-degrading bacteria produced a glycolipid biosurfactant in the process of fermentation and degradation, which can promote the solubilization, emulsification, wetting and penetration of long-chain alkanes, and can effectively degrade phenol in coking wastewater and petrochemical wastewater.

The present invention discloses a multiple PCR detection kit for rapidly identifying pathogenic Pseudomonas plecoglossicida, and a method thereof. The kit comprises TaqDNA polymerase, a PCR buffer, a deoxyribonucleoside triphosphate mixture, double distilled water, and primer pairs of Pseudomonadaceae, Pseudomonas plecoglossicida and pathogenic Pseudomonas plecoglossicida. The multiple PCR detection method for rapidly identifying pathogenic Pseudomonas plecoglossicida by using the kit comprises: primary separation of Pseudomonadaceae, and multiple PCR amplifications. Compared with the kit and the method in the prior art, the kit and the method of the present invention have the following advantages that: with application of the kit of the present invention to detect, Pseudomonadaceae, Pseudomonas plecoglossicida and pathogenic Pseudomonas plecoglossicida corresponding to the DNA primer pairs of the three Pseudomonadaceae can be rapidly detected, and with the detection method of the present invention, the presence of the Pseudomonadaceae can be rapidly, sensitively and specifically determined by using the kit, and the pathogenic stain and the non-virulent environment isolation strain can be distinguished.

The invention relates to construction of a recombinant strain capable of producing arginine deiminase (ADI) and a directional modification method thereof, belonging to the technical field of medical biological engineering. The method comprises the following steps of: amplifying the ADI coding gene arcA of a pseudomonas plecoglossicida CGMCC (China General Microbiological Culture Collection Center) No. 2039 by adopting a PCR (polymerase chain reaction) method, and constructing an ADI recombination expression strain, researching the enzymology properties of the recombinant ADI enzyme, wherein Km is 2.88mmol / L (pH is 6.0), the optimal pH is 6.0, the enzyme activity is 20.85U / mg, and the enzyme activity is reduced by more than 90% when the pH is increased to the physiological pH (7.4). The directional modification is carried out on the recombinant ADI enzyme by adopting a prone PCR mutation technology, so as to improve the activity and substrate affinity of the enzyme under the physiological pH condition. An excellent mutant strain ADIM314 is obtained by screening through the directional modification. Compared with a wild enzyme, the activity of the ADIM314 enzyme under the physiological pH condition is improved by more than 20 times, the Km value is reduced to 0.65mmol / L (pH is 7.4), and the optimal pH is improved to 6.5 from 6.0.