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Extraction method of mycelial fungus genome

A technology of filamentous fungus and extraction method, which is applied in the field of genome extraction of Penicillium chrysogenum, can solve the problems of corrosiveness, toxicity, and lengthy extraction process of organic reagents, and achieve the goal of reducing damage, simplifying the operation process, and avoiding the use Effect

Inactive Publication Date: 2015-09-30
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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Problems solved by technology

These methods can produce higher-quality genomic DNA, which can meet the subsequent genetic manipulation requirements such as PCR reactions. However, for the molecular identification and screening of tens of thousands of filamentous fungal transformants, the application of these conventional methods to extract genomes is difficult. The complicated extraction process is an important link that limits the progress of the experiment; in addition, in the conventional extraction method, various reagents are used to break the cell wall, phenol / chloroform extraction to remove protein, ethanol or isopropanol precipitation and other multi-step extractions. DNA, the extraction process is lengthy, and the organic reagent is corrosive and toxic, which is not good for the health of the operator

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  • Extraction method of mycelial fungus genome
  • Extraction method of mycelial fungus genome
  • Extraction method of mycelial fungus genome

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Embodiment 1

[0033] This example relates to the extraction method of filamentous fungal genome.

Embodiment 11

[0035] This embodiment relates to a method for extracting genomes of filamentous fungi.

Embodiment 111

[0037] This embodiment relates to a method for extracting the genome of Penicillium chrysogenum, comprising the following extraction steps:

[0038] a. Obtain the mycelia of Penicillium chrysogenum: inoculate the rice spores of Penicillium chrysogenum on solid medium for culture, pick the clone to obtain the mycelium of Penicillium chrysogenum; 3 5g / L Penicillium chrysogenum liquid growth medium, shake culture at 26°C for 48h, centrifuge at 10000rpm for 2min to obtain the mycelia of Penicillium chrysogenum;

[0039] b. Add enzyme solution: add 60 μL of helicase solution containing helicase concentration of 10 mg / mL and 40 μL of cellulase solution containing cellulase concentration of 10000 U / mL to the mycelia of Penicillium chrysogenum in step a, totaling 100 μL of mixed enzyme solution , to obtain the Penicillium chrysogenum mycelia liquid; the mixed enzyme liquid is prepared by dissolving commercially available helicase and cellulase in the penetrant respectively, the helica...

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Abstract

The invention relates to an extraction method of a mycelial fungus genome. Enzymolysis is performed on Penicillium chrysogenum, Aspergillus niger, Trichoderma reesei, penicillium citrinum and other mycelial fungus hyphae by using a helicase-cellulase mixed enzyme solution, and the enzymolysis treatment is effectively combined with ultrasonic treatment and high-temperature heating treatment, thereby obtaining the mycelial-fungus-genome-containing supernate for PCR (polymerase chain reaction) identification. The supernate of the mycelial fungus hypha enzymolysis solution obtained by the extraction method is directly used for PCR reaction to obtain the mycelial fungus 18S rDNA gene strip without purification operation. The operational process is simple, safe and stable, avoids the use of the toxic extraction reagent, and reduces the injury to the experimental personnel.

Description

technical field [0001] The invention relates to a genome extraction method of filamentous fungi, in particular to a genome extraction method of Penicillium chrysogenum. Background technique [0002] Among industrial microorganisms, filamentous fungi are a very important group, especially in the three major production fields of antibiotics, organic acids, and enzyme preparations. The production performance of filamentous fungi is extremely outstanding, such as Penicillium chrysogenum ( Penicillium chrysogenum ) is an important penicillin-producing filamentous fungus; Aspergillus niger ( Aspergillus niger ) is an important industrial fermentation strain of citric acid; Trichoderma reesei ( Trichoderma reesei ) is an important industrial fermentation strain of cellulase. In order to further increase the production of penicillin, citric acid and cellulase, the transformation of industrial strains at the molecular level has broad application prospects. The extraction of...

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Application Information

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IPC IPC(8): C12N15/10C12R1/82C12R1/685C12R1/885C12R1/80
Inventor 张春晓韩梁彦王丽丽
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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