Method for inserting gene of pseudomonas fluorescens M18 into mutation strain H18G

A Pseudomonas fluorescens, insertion mutation technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problem of low growth rate, achieve the effect of increased synthesis capacity and broad application prospects

Inactive Publication Date: 2004-10-27
SHANGHAI JIAO TONG UNIV
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  • Method for inserting gene of pseudomonas fluorescens M18 into mutation strain H18G
  • Method for inserting gene of pseudomonas fluorescens M18 into mutation strain H18G

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[0014] Further describe embodiment below in conjunction with technical scheme of the present invention

[0015] 1. Under the improved PPM medium fermentation conditions, compared with the wild strain M18, the ability of the mutant strain M18G to synthesize phenazine-1-carboxylic acid was increased by 2 times. The mutant strain M18G and the wild strain M18 were respectively inoculated into Erlenmeyer flasks containing 150ml of PPM medium. The volume of the Erlenmeyer flasks was 500ml, and placed in a shaker at 28°C and 220r / min for fermentation and culture, and samples were taken every 8 hours Analysis was repeated three times and the average value was taken. After 72 hours of fermentation, the HPLC measurement results show that the amount of PCA biosynthesized by M18G can reach 121.5ppm, and the amount of PCA biosynthesized by M18 is only 60.6ppm, see figure 1 .

[0016] 2. Under the fermentation conditions of King'B medium, compared with the wild strain M18, the ability of...

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Abstract

A method for inserting the gene of fluorescent pseudomonads M18 in the mutant strain M18G in order to develop efficient low-poison agricultural chemical includes designing facultative primer, PCR amplification of gacA fragment in M18 genon, using it as probe to screen positive clone from the plasmid library of M18 genom, using homogeneous recombination reject technique and solid-phase filter membrane triparent cross to obtain mutant strain M18G, and extracting and quantitatively testing phenazine-1-carboxylate.

Description

technical field [0001] The invention relates to a method for gene insertion mutation strain of Pseudomonas, in particular to a method for gene insertion mutation strain M18G of Pseudomonas fluorescens M18, which belongs to the field of gene technology. Background technique [0002] It is generally believed that the yield reduction caused by crop diseases accounts for about 25-75% of the total income. At present, in addition to the selection of improved varieties and scientific cultivation measures, the main means of controlling crop diseases in production is the use of chemical fungicides. Most fungicides are harmful to animals and humans. Food poisoning caused by chemical pesticides occurs from time to time. The potential harm of poisons left in edible parts to humans and animals has attracted the attention of all aspects of society. The long-term accumulation of chemical pesticides in the ecosystem has caused environmental pollution, which is extremely unfavorable to the ...

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Application Information

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IPC IPC(8): C12N1/21C12N15/31C12N15/63C12P1/04C12Q1/68
Inventor 许煜泉葛宜和
Owner SHANGHAI JIAO TONG UNIV
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