ArticleArchaea (Vancouver, B.C.)2011
Establishing a markerless genetic exchange system for Methanosarcina mazei strain Gö1 for constructing chromosomal mutants of small RNA genes.
Article in Archaea (Vancouver, B.C.), 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 27 citations in OpenAlex.
- 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
- CRISPR/Cas12a toolbox for genome editing inFrontiers in microbiology · 2023Article
- Overview of Diverse Methyl/Alkyl-Coenzyme M Reductases and Considerations for Their Potential Heterologous Expression.Frontiers in microbiology · 2022Review
- Genetic Methods and Construction of Chromosomal Mutations in Methanogenic Archaea.Methods in molecular biology (Clifton, N.J.) · 2022Article
- sRNARNA biology · 2017Article
- The apt/6-Methylpurine Counterselection System and Its Applications in Genetic Studies of the Hyperthermophilic Archaeon Sulfolobus islandicus.Applied and environmental microbiology · 2016Article
- Small regulatory RNAs in Archaea.RNA biology · 2014Review
- Generation and phenotyping of a collection of sRNA gene deletion mutants of the haloarchaeon Haloferax volcanii.PloS one · 2014Article
- Contribution of transcriptomics to systems-level understanding of methanogenic Archaea.Archaea (Vancouver, B.C.) · 2013Review
- A novel inducible protein production system and neomycin resistance as selection marker for Methanosarcina mazei.Archaea (Vancouver, B.C.) · 2012Article
- Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A markerless genetic exchange system was successfully established in Methanosarcina mazei strain Gö1 using the hpt gene coding for hypoxanthine phosphoribosyltransferase. First, a chromosomal deletion mutant of the hpt gene was generated conferring resistance to the purine analog 8-aza-2,6-diaminopurine (8-ADP). The nonreplicating allelic exchange vector (pRS345) carrying the pac-resistance cassette for direct selection of chromosomal integration, and the hpt gene for counterselection was introduced into this strain. By a pop-in and ultimately pop-out event of the plasmid from the chromosome, allelic exchange is enabled. Using this system, we successfully generated a M. mazei deletion mutant of the gene encoding the regulatory non-coding RNA sRNA(154). Characterizing M. mazei ΔsRNA(154) under nitrogen limiting conditions demonstrated differential expression of at least three cytoplasmic proteins and reduced growth strongly arguing for a prominent role of sRNA(154) in regulation of nitrogen fixation by posttranscriptional regulation.
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Registered trials
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.