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Filamentous fungus replicon

A filamentous fungus and replicon technology, applied in biochemical equipment and methods, fermentation, plant gene improvement, etc., can solve the problems of low efficiency of integration and transformation, lack of plasmids, etc., and achieve rich component library, high stability, and expansion The effect of choosing diversity

Active Publication Date: 2022-08-09
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Before the discovery of the fungal autonomous replication sequence, the main defect of the filamentous fungal transformation system is that there is no plasmid similar to that that can replicate autonomously outside the chromosome of yeast, resulting in low efficiency of integration and transformation

Method used

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Embodiment Construction

[0036] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.

[0037] 1 Fungal culture and genome extraction

[0038] 1.1 Fungal culture

[0039] Take the strain out of the -80 ℃ refrigerator, thaw naturally on ice, dip the bacterial liquid in the inoculation loop burned by the alcohol lamp in the ultra-clean workbench, streak the solid PDA medium, and darken the culture conditions according to Table 1. Cultivate until a single colony grows, pick a part of the bacterial cells and mix them in sterile water, take 200 μL of the spore suspension and cultivate it in the PDA solid medium covered with cellophane until the medium is covered with hyphae, and refrigerate at 4°C. save.

[0040] Table 1 Fungal species and culture conditions

[0041]

[0042] 1.2 Fungal genome extraction method

[0043] The cultured fungal mycelium was used as the template to extract the genome, and the specific steps we...

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Abstract

The invention discloses a filamentous fungus replicon. The nucleotide sequence of the replicon is as shown in SEQ ID NO. 2. The replicon is obtained by using 19 fungal genomes as templates to construct a metagenome library and transforming aspergillus niger spores for screening, and is from a rhizopus chlamydosporum endophyte burkholderia HKI454 genome. The replicon provided by the invention can play a replication function in an aspergillus niger expression system, can provide stability higher than that of an AMA1 replicon, is a replicon found in fungi except aspergillus nidulans for the first time, enriches a replicon element library, and expands selection diversity of filamentous fungi utilizing the replicon.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a filamentous fungal replicon. Background technique [0002] Before the fungal autonomous replication sequence was discovered, the main defect of the filamentous fungal transformation system was that there was no plasmid that could replicate autonomously outside the chromosome of yeast, resulting in low efficiency of integrated transformation. Since researchers discovered the ARS sequence in Saccharomyces cerevisiae, research on replicons has been carried out in fungi. A 6.1Kb sequence AMA1 was first discovered in Aspergillus nidulans. The plasmid containing the AMA1 sequence can be transformed into Aspergillus nidulans to improve the transformation efficiency 250 times, and can replicate autonomously outside the chromosome, the sequence has also been applied to other fungi, such as Penicillium chrysogenum, Aspergillus niger, Aspergillus parasiticus, etc. Since the discovery of the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/55C12N15/80C12N15/66C12N15/64
CPCC07K14/195C12N9/14C12N15/80C12Y306/04012C12N2820/007C12N2820/55Y02A50/30
Inventor 林峻燕天鹤
Owner FUZHOU UNIV
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