Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Nucleotide specific to citrobacter O6 and O9 as well as application of nucleotide

A technology of Citrobacter and nucleotides, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of lack of previous research foundation, unpredictable gene function, and difficulty in identification of sugar synthesis genes. And other issues

Active Publication Date: 2016-01-06
NANKAI UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reasons for the above phenomenon are: polysaccharide antigen synthesis gene cluster composition is complex, gene function is difficult to predict, sugar synthesis gene identification is difficult, etc.
In addition, most of the units that have carried out some relevant research lack the foundation of previous research

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Nucleotide specific to citrobacter O6 and O9 as well as application of nucleotide
  • Nucleotide specific to citrobacter O6 and O9 as well as application of nucleotide
  • Nucleotide specific to citrobacter O6 and O9 as well as application of nucleotide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 : Genome Extraction

[0033] Citrobacter was cultured at 37°C in brain-heart infusion broth medium, the bacteria were collected, and the genome was extracted with a bacterial genome extraction kit (Kangwei Century CW0552). The specific steps are as follows:

[0034] 1. Take 1-5ml of bacterial culture (106-108 cells, no more than 2×109 cells), centrifuge at 10000rpm for 1 minute, and discard the supernatant.

[0035] 2. Add 180μl BufferGTL to the pellet and shake to resuspend the bacteria.

[0036] 3. Add 20 μl ProteinaseK, vortex to mix, and incubate at 56°C until the cells are completely lysed. During the incubation, invert or shake the centrifuge tube at regular intervals to disperse the sample.

[0037] 4. Add 4 μl of RNaseA solution (Product No. CW0601) with a concentration of 100 mg / ml, vortex to mix, incubate at room temperature for 5 minutes, and then proceed to step 5.

[0038] 5. Add 200μl BufferGL, vortex and mix well; add 200μl absolute ethanol,...

Embodiment 2

[0050] Example 2: sequence deciphering

[0051] The genomes of the standard strains of Citrobacter serotype O6 and O9 were extracted, and the genomes of each serotype of Citrobacter were sequenced by Solexapair-end sequencing technology to obtain the sequence of the serotype, and the sequences were compared by using Blast and PSI-Blast. The TMHMM2.0 program was used to predict the transmembrane structure, and the ClustalW program was used to perform sequence alignment and screen conservative and specific gene fragments. Finally, the O antigen gene cluster sequences and decipher results of each serotype of Citrobacter were obtained.

Embodiment 3

[0052] Example 3 : Primer design

[0053] According to the deciphering of gene clusters, we found that wxya The gene is indeed a serotype-specific gene, so a specific segment of the gene is selected to design specific primers.

[0054] Primer design is a core part of this invention. Primers were designed according to the specific genes described in the literature. wxya The gene is a relatively specific gene in the Citrobacter O antigen gene cluster, which can be used as the target gene for serotype identification. Import the above genes into PrimerPremier5 for primer design. The length of the primers should preferably be between 18 and 24 bp, and the Tm value should be around 55°C. A pair of primers are designed for each gene, with a single target band.

[0055] After the primers are designed, BLAST is performed in Genbank. The designed primers should not have too high sequence similarity with other related bacteria, so as to ensure that the primers can only amplify ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a nucleotide specific to citrobacter O6 and O9 as well as an application of the nucleotide. The nucleotide is at least one of nucleotides shown in SEQ ID NO:1-4. Those nucleotides can be applied to preparation of a PCR (polymerase chain reaction) kit for detecting citrobacter. The citrobacter can be sampled from digestive attracts, urine, blood and crude extracts of wound cultures and can be separated from human and animal waste environment. According to the nucleotide specific to citrobacter O6 and O9 serotypes as well as the PCR kit comprising the nucleotide, the PCR kit is convenient to prepare, short in detection period, high in speed, high in operability, prone to industrial production, lower in detection cost and high in accuracy and sensitivity.

Description

technical field [0001] The present invention relates to nucleotides specific to Citrobacter O6 and O9 serotypes, in particular to nucleotides specific to a single gene in the O antigen gene cluster of Citrobacter O6 and O9 serotypes and application thereof. Background technique [0002] Citrobacter is a common Gram-negative bacterium in the Enterobacteriaceae family, which can usually use citric acid as the only carbon source and belongs to facultative anaerobic bacteria. Citrobacter, the closest relative to Salmonella and Escherichia coli, is an opportunistic pathogen. [0003] Citrobacter can be divided into 11 gene species by DNA hybridization method, namely Citrobacter freundii (C.freundii), Citrobacter koseri (C. koseri), Citrobacter nonmalonate (C. .amalonatIzcus), Citrobacter fageri (C.farmeri), Citrobacter youngae (C.youngae), Citrobacter braakii (C.braakii), Citrobacter walkmanii (C.werkmanii), plug C. sedlakif, C. rodentium, C. gillenii, C. murliniae, which have ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68C12Q1/04
Inventor 王磊钱澄倩郭玺刘斌
Owner NANKAI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com