ArticleCurrent genetics2005
A two-step protocol for efficient deletion of genes in the filamentous ascomycete Podospora anserina.
Article in Current genetics, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 30 citations in OpenAlex.
- Simultaneous Ablation of the Catalytic AMPK α-Subunit SNF1 and Mitochondrial Matrix Protease CLPP Results in Pronounced Lifespan Extension.Frontiers in cell and developmental biology · 2021Article
- Impact of F1Fo-ATP-synthase dimer assembly factors on mitochondrial function and organismic aging.Microbial cell (Graz, Austria) · 2018Article
- Cyclophilin D Is Involved in the Regulation of Autophagy and Affects the Lifespan of P. anserina in Response to Mitochondrial Oxidative Stress.Frontiers in genetics · 2016Article
- Identification of potential mitochondrial CLPXP protease interactors and substrates suggests its central role in energy metabolism.Scientific reports · 2015Article
- RCF1-dependent respiratory supercomplexes are integral for lifespan-maintenance in a fungal ageing model.Scientific reports · 2015Article
- Overexpression of Pa_1_10620 encoding a mitochondrial Podospora anserina protein with homology to superoxide dismutases and ribosomal proteins leads to lifespan extension.Current genetics · 2015Article
- Identification of autophagy as a longevity-assurance mechanism in the aging model Podospora anserina.Autophagy · 2014Article
- Poly(ADP-ribose) polymerase is a substrate recognized by two metacaspases of Podospora anserina.Eukaryotic cell · 2013Article
- Human CLPP reverts the longevity phenotype of a fungal ClpP deletion strain.Nature communications · 2013Article
- Unmasking a temperature-dependent effect of the P. anserina i-AAA protease on aging and development.Cell cycle (Georgetown, Tex.) · 2011Article
- PaCATB, a secreted catalase protecting Podospora anserina against exogenous oxidative stress.Aging · 2011Article
- Modulation of the glyoxalase system in the aging model Podospora anserina: effects on growth and lifespan.Aging · 2010Article
- Deletion of PaAif2 and PaAmid2, two genes encoding mitochondrial AIF-like oxidoreductases of Podospora anserina, leads to increased stress tolerance and lifespan extension.Current genetics · 2010Article
- Increasing organismal healthspan by enhancing mitochondrial protein quality control.Nature cell biology · 2009Article
- Age-related cellular copper dynamics in the fungal ageing model Podospora anserina and in ageing human fibroblasts.PloS one · 2009Article
- Gene deletion and allelic replacement in the filamentous fungus Podospora anserina.Current genetics · 2008Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Deletion of genes in Podospora anserina via conventional methods is an inefficient and time-consuming process since homologous recombination occurs normally only at low frequency (about 1%). To improve the efficiency of replacement, we adopted the two-step protocol developed for Aspergillus nidulans (Chaveroche et al. in Nucleic Acids Res 28:E97, 2000). As a prerequisite, a vector was generated containing a blasticidin resistance cassette for selection in the Escherichia coli host strain KS272 (pKOBEG) and a phleomycin resistance cassette for selection in P. anserina. A derivative of this vector, into which short ( approximately 250 bp) PCR-generated sequences flanking the gene to be deleted have been integrated, is introduced into the E. coli host strain which contains a cosmid with the gene of interest and long 5' and 3' flanking sequences. Subsequently, a cosmid is reisolated from E. coli in which the gene of interest is replaced by the resistance cassette. This construct is used to transform P. anserina. The long stretches flanking the resistance cassette facilitate recombination with homologous sequences in the fungal genome and increase the efficiency of gene deletion up to 100%. The procedure is not dependent on the availability of specific auxotrophic mutant strains and may be applicable to other fungi.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.