ArticleMolecular genetics and genomics : MGG2005
Development of genetic methods and construction of a chromosomal glnK1 mutant in Methanosarcina mazei strain Gö1.
Article in Molecular genetics and genomics : MGG, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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28 citing papers in PubMed, 56 citations in OpenAlex.
- Article
- HThe FEBS journal · 2026Article
- Structure of the Methanosarcina mazei Mtr complex bound to the oxygen-stress responsive small protein MtrI.Nature communications · 2025Article
- Production of the Sesquiterpene Bisabolene From One- and Two-Carbon Compounds in Engineered Methanosarcina acetivorans.Microbial biotechnology · 2025Article
- Functional insights into Solo-Cas4 inmicroLife · 2025Article
- Uncovering the small proteome of Methanosarcina mazei using Ribo-seq and peptidomics under different nitrogen conditions.Nature communications · 2024Article
- Teaching old dogs new tricks: genetic engineering methanogens.Applied and environmental microbiology · 2024Review
- Compiling a versatile toolbox for inducible gene expression inmicroLife · 2024Article
- Active in vivo translocation of the Methanosarcina mazei Gö1 Casposon.Nucleic acids research · 2023Article
- Newly Established Genetic System for Functional Analysis of MetSV.International journal of molecular sciences · 2023Article
- Generating a Small Shuttle Vector for Effective Genetic Engineering ofInternational journal of molecular sciences · 2022Article
- Insights into the biotechnology potential ofFrontiers in microbiology · 2022Review
- Overview of Diverse Methyl/Alkyl-Coenzyme M Reductases and Considerations for Their Potential Heterologous Expression.Frontiers in microbiology · 2022Review
- The CARF Protein MM_0565 Affects Transcription of the Casposon-EncodedBiomolecules · 2020Article
- Archaeosine Modification of Archaeal tRNA: Role in Structural Stabilization.Journal of bacteriology · 2020Article
- Cross-cleavage activity of Cas6b in crRNA processing of two different CRISPR-Cas systems in Methanosarcina mazei Gö1.RNA biology · 2019Article
- sRNARNA biology · 2017Article
- Small regulatory RNAs in Archaea.RNA biology · 2014Review
- Two CRISPR-Cas systems in Methanosarcina mazei strain Gö1 display common processing features despite belonging to different types I and III.RNA biology · 2013Article
- An archaeal sRNA targeting cis- and trans-encoded mRNAs via two distinct domains.Nucleic acids research · 2012Article
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Authors and funding
6 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The methanogenic archaeon Methanosarcina mazei strain Gö1 has so far proven to be genetically intractable due to its low plating efficiency on solid medium and the lack of an effective transformation method. Here, we report the first significant improvement in plating efficiency (up to 10%), which was achieved by (1) selecting for a spontaneous mutant of M. mazei that shows significantly higher resistance to mechanical stress during spreading an agar plates, and (2) plating the cells in 0.5% top agar with trimethylamine as a carbon and energy source under a H2S-containing atmosphere (0.1%). Using this mutant we succeeded in establishing a liposome-mediated transformation protocol, which for the first time allowed genetic manipulation of the M. mazei Gö1 strain. We further report on the construction of the first chromosomal deletion mutant of M. mazei by means of homologous recombination. Characterization of this mutant strain revealed that M. mazei cells lacking a functional glnK1-gene exhibited a partial growth defect under nitrogen limitation when molecular nitrogen was used as the sole nitrogen source. Quantitative RT-PCR analysis, however, showed that genes involved in nitrogen assimilation or nitrogen fixation are transcribed in the glnK1 mutant as in the wild type. Thus, we propose that the archaeal GlnK1 protein is not directly involved in the transcriptional regulation of genes involved in nitrogen metabolism, but rather affects their protein products directly.
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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.