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Regulators involved in mushroom formation

A technology for mushrooms and fungi, applied in the field of regulators involved in the formation of mushrooms, can solve problems such as genes not being described

Inactive Publication Date: 2012-05-16
UTRECHT UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Genes involved in the regulation / initiation of mushroom production have not been described so far

Method used

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  • Regulators involved in mushroom formation
  • Regulators involved in mushroom formation
  • Regulators involved in mushroom formation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0129] Example 1: Confirmation of hypothetical transcription factors in the Schizophyllum genome

[0130] passJoint Genome Institute automatically called genes of Schizophyllum strain 4-8 (FGSC#9210) genome (8.29X coverage) and obtained 13181 predicted genes (see http: / / jgi.doe.gov / Scommune). These genes were automatically annotated using GO (Gene Ontology) (Ashburner et al., 2000), KOG (euKaryotic Orthologous Groups) (Koonin et al., 2004) and PFAM algorithm (Finn et al., 2008). This automated annotation serves as a starting point to identify transcription factors (TFs):

[0131] • 190 genes were placed in the GO entry "Transcription Factor Activity" based on known DNA binding domains (GO:0003700).

[0132] · The KOG annotation algorithm predicted 569 genes commonly involved in transcription (Functional ID: "K"). Of these, 205 transcription factors were manually identified.

[0133] • Based on the presence of the PFAM domain PF00096 / IPR007087 (zinc finger structure, C2H...

Embodiment 2

[0173] Example 2: Proof of principle: Knockout of the fst3 (protein ID 257422) gene affects mushroom development

[0174] The putative transcription factor gene fst3 (Protein ID 257422) was knocked out. For this, the pDelcas vector described in Ohmetal. (2010) was used. Primers used to create knockout constructs are shown in Table 2. This knockout construct, designated pRO097, contains regions flanking the coding sequence of fst3 with the nourthricin resistance gene in between. The phleomycin resistance gene cassette is located elsewhere in this construct (see Ohm et al., 2010 for details). Schizophyllum strain H4-8 was modified as described by van Peer et al., 2009. Regeneration medium does not contain antibiotics, where selection plates contain 20 μg ml -1 Nourthricin. The deletion of the target gene was verified by PCR (see Ohm et al., 2010 for the procedure). Monokaryons compatible with the gene deletion were selected from spores produced by crossing the mutant str...

Embodiment 3

[0175] Example 3: Proof of principle: Knockout of fst4 (Protein ID 66861) affects mushroom development.

[0176] The putative transcription factor gene fst4 (Protein ID 66861) was knocked out. For this, the pDelcas vector described in Ohmetal. (2010) was used. Primers used to create knockout constructs are shown in Table 2. This knockout construct, designated pRO191, contains regions flanking the coding sequence of fst4 with the nourthricin resistance gene in between. The phleomycin resistance gene cassette is located elsewhere in this construct (see Ohm et al., 2010 for details). Schizophyllum strain H4-8 was modified as described by van Peer et al., 2009. Regeneration medium does not contain antibiotics, where selection plates contain 20 μg ml -1 Nourthricin. The deletion of the target gene was verified by PCR (see Ohm et al., 2010 for the procedure). Monokaryons compatible with the gene deletion were selected from spores produced by crossing the mutant strain with t...

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Abstract

The invention relates to a fungus or a mushroom and to a method of producing it wherein the fungus / mushroom has an increased expression level of a polypeptide and / or has a decreased expression level of a polypeptide, wherein the polypeptide comprises an amino acid sequence that has at least 40% amino acid identity or similarity with a sequence selected from SEQ ID NO:1-200 and / or that has at least 50% amino acid identity or similarity with a sequence selected from SEQ ID NO:201-208.

Description

technical field [0001] The present invention relates to mushrooms and methods for producing said mushrooms, wherein the mushrooms have increased expression levels of polypeptides and / or reduced expression levels of polypeptides, wherein the polypeptides are encoded by an amino acid sequence selected from SEQ ID NO: 1-200 having A protein with at least 40% identity or similarity and / or a protein with at least 50% identity or similarity to an amino acid sequence selected from SEQ ID NO: 201-208 is encoded by a nucleotide sequence. Background technique [0002] Mushroom formation is a highly complex developmental process. As an example, we describe a general protocol for fruiting body formation of agarics such as Agaricus bisporus (see Kües, 2000; Umar and van Griensven, 1997). After the formation of a "critical mass" of submerged mycelium, the mycelium breaks away from the medium and grows into the air. These hyphae form aggregates, so-called hyphal nodules or hyphal nodules...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/375
CPCC07K14/375
Inventor R·A·奥姆J·F·迪琼L·G·卢格尼斯H·A·B·沃斯顿
Owner UTRECHT UNIVERSITY
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