Preparation and detection method of pseudomonas aeruginosa algC gene clonal strains

A Pseudomonas aeruginosa detection method technology, applied in the field of animal genetic engineering, can solve the problem of few research reports

Inactive Publication Date: 2015-02-25
GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Scholars at home and abroad have also found that human alginate antibodies have the effect of preventing and treating bacterial biofilms [Mai GT, et al (1993)] and alginate immunogenicity and immunological functions [Chai Dong, et al (2004); Li Hongyan, etc. (2005), and studied the biology and function of the regulatory genes algX, algE, alg44 [J.Gutsche, et al, (2006); Uwe Remminghorst, et al (2006); Yang Maosheng, et al (2008)] , but there are few reports on algC synthesis gene algC, which encodes phosphomannitol mutase and consists of 1392 base pairs

Method used

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  • Preparation and detection method of pseudomonas aeruginosa algC gene clonal strains
  • Preparation and detection method of pseudomonas aeruginosa algC gene clonal strains
  • Preparation and detection method of pseudomonas aeruginosa algC gene clonal strains

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Embodiment Construction

[0030] Embodiments of the present invention: a preparation and detection method of Pseudomonas aeruginosa algC gene clone:

[0031] 1 Strains and culture methods

[0032]In this experiment, Pseudomonas aeruginosa was isolated from goats and wild animal bamboo rat disease materials. The medium for cultivating Pseudomonas aeruginosa was LB medium, PIA medium, GXI medium, SGI medium, and MSM medium. After the strain was cultured at 37 °C for about 24 h-72 h, 1 ml of the culture solution was taken and transferred to a container. In a preservation tube with 70 μL DMSO, the strains were stored at -70°C for future use. NTP, Taq, Pfx enzymes, and E.coli TOP10 were provided by Invitrogen. E. coli TOP10 was used to transform Escherichia coli containing the target gene plasmid. When cultivating the strains, add appropriate amount of antibiotics, such as ampicillin, 100 μg / ml; Amamicin, 10 μg / ml; tetracycline, 12.5 μg / ml; plasmid pGEM-TEasy provided by Promega.

[0033] 2 Preparation o...

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Abstract

The invention discloses a preparation method of clonal strains containing pseudomonas aeruginosa algC gene. The method comprises the following steps: culturing bacterial strain DNA, preparing chromosome, preparing plasmid DNA, optimizing PCR, purifying PCR products, cloning algC gene and preserving clonal strains. The invention also discloses a detection method of clonal strains containing pseudomonas aeruginosa algC gene. The invention provides materials and technique for the research and development of pseudomonas aeruginosa alginate structural gene and alginate products and provides experimental materials, techniques and molecular biological basis theory for environmental safety evaluation test.

Description

technical field [0001] The invention relates to the field of animal genetic engineering, in particular to a method for preparing and detecting a Pseudomonas aeruginosa algC gene clone. Background technique [0002] Pseudomonas aeruginosa was first isolated from wound pus by Gessard in 1882, and it is one of the more common clinical opportunistic pathogens. Pseudomonas aeruginosa, as an opportunistic pathogen, has also been reported to infect animals such as poultry, cattle, sheep, rabbits, giant pandas, ducks, quails, roe deer, and gibbons. Human Pseudomonas aeruginosa, which attacks the lungs of patients with cystic fibrosis, is a drug-resistant variant that forms biofilms and is associated with pathogenic alginate formation, research suggests. Scholars at home and abroad have also found that human alginate antibodies have the effect of preventing and treating bacterial biofilms [Mai GT, et al (1993)] and alginate immunogenicity and immunological functions [Chai Dong, et a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12Q1/68C12Q1/04C12R1/385
Inventor 杨茂生杨莉龙明珠
Owner GUIZHOU INST OF ANIMAL HUSBANDRY & VETERINARY
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