ArticleArchaea (Vancouver, B.C.)2012
A novel inducible protein production system and neomycin resistance as selection marker for Methanosarcina mazei.
Article in Archaea (Vancouver, B.C.), 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 16 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
16 citing papers in PubMed, 20 citations in OpenAlex.
- Valine overproduction inMetabolic engineering communications · 2026Article
- Distinct Patterns of Antibiotic Sensitivities in Ammonia-Oxidising Archaea.Environmental microbiology · 2025Article
- Teaching old dogs new tricks: genetic engineering methanogens.Applied and environmental microbiology · 2024Review
- Probing Denitrifying Anaerobic Methane Oxidation via Antimicrobial Intervention: Implications for Innovative Wastewater Management.Environmental science & technology · 2024Article
- Compiling a versatile toolbox for inducible gene expression inmicroLife · 2024Article
- Newly Established Genetic System for Functional Analysis of MetSV.International journal of molecular sciences · 2023Article
- Application of the Fluorescence-Activating and Absorption-Shifting Tag (FAST) for Flow Cytometry in Methanogenic Archaea.Applied and environmental microbiology · 2023Article
- Editorial: The methane moment - Cross-boundary significance of methanogens: Preface.Frontiers in microbiology · 2022Article
- 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
- The streptothricin acetyltransferase (sat) gene as a positive selectable marker for methanogenic archaea.FEMS microbiology letters · 2019Article
- Biotechnology of extremely thermophilic archaea.FEMS microbiology reviews · 2018Review
- Methanogens: biochemical background and biotechnological applications.AMB Express · 2018Review
- sRNARNA biology · 2017Article
- Development of β -lactamase as a tool for monitoring conditional gene expression by a tetracycline-riboswitch in Methanosarcina acetivorans.Archaea (Vancouver, B.C.) · 2014Article
- Article
Corrections and comments
- Erratum issued
Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Methanosarcina mazei is one of the model organisms for the methanogenic order Methanosarcinales whose metabolism has been studied in detail. However, the genetic toolbox is still limited. This study was aimed at widening the scope of utilizable methods in this group of organisms. (i) Proteins specific to methanogens are oftentimes difficult to produce in E. coli. However, a protein production system is not available for methanogens. Here we present an inducible system to produce Strep-tagged proteins in Ms. mazei. The promoter p1687, which directs the transcription of methyl transferases that demethylate methylamines, was cloned into plasmid pWM321 and its activity was determined by monitoring β-glucuronidase production. The promoter was inactive during growth on methanol but was rapidly activated when trimethylamine was added to the medium. The gene encoding the β-glucuronidase from E. coli was fused to a Strep-tag and was cloned downstream of the p1687 promoter. The protein was overproduced in Ms. mazei and was purified in an active form by affinity chromatography. (ii) Puromycin is currently the only antibiotic used as a selectable marker in Ms. mazei and its relatives. We established neomycin resistance as a second selectable marker by designing a plasmid that confers neomycin resistance in Ms. mazei.
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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.