ArticleNucleic acids research2005
A gene truncation strategy generating N- and C-terminal deletion variants of proteins for functional studies: mapping of the Sec1p binding domain in yeast Mso1p by a Mu in vitro transposition-based approach.
Article in Nucleic acids research, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed.
- Alkaliphilic microbial cellulases: sources, structural insights, and bioengineering approaches for industrial applications.Archives of microbiology · 2026Review
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- Iterative screening methodology enables isolation of strains with improved properties for a FACS-based screen and increased L-DOPA production.Scientific reports · 2019Article
- BpeB, a major resistance-nodulation-cell division transporter from Burkholderia cenocepacia: construct design, crystallization and preliminary structural analysis.Acta crystallographica. Section F, Structural biology communications · 2018Article
- Extensive libraries of gene truncation variants generated by in vitro transposition.Nucleic acids research · 2017Article
- MuA-mediated in vitro cloning of circular DNA: transpositional autointegration and the effect of MuB.Molecular genetics and genomics : MGG · 2016Article
- Generation of comprehensive transposon insertion mutant library for the model archaeon, Haloferax volcanii, and its use for gene discovery.BMC biology · 2014Article
- An assay to monitor the activity of DNA transposition complexes yields a general quality control measure for transpositional recombination reactions.Mobile genetic elements · 2014Article
- A 'resource allocator' for transcription based on a highly fragmented T7 RNA polymerase.Molecular systems biology · 2014Article
- A transposase strategy for creating libraries of circularly permuted proteins.Nucleic acids research · 2012Article
- Article
- Universal platform for quantitative analysis of DNA transposition.Mobile DNA · 2010Article
- Mso1p regulates membrane fusion through interactions with the putative N-peptide-binding area in Sec1p domain 1.Molecular biology of the cell · 2010Article
- Transposition-based method for the rapid generation of gene-targeting vectors to produce Cre/Flp-modifiable conditional knock-out mice.PloS one · 2009Article
- Bacteriophage Mu integration in yeast and mammalian genomes.Nucleic acids research · 2008Article
- High-precision mapping of protein protein interfaces: an integrated genetic strategy combining en masse mutagenesis and DNA-level parallel analysis on a yeast two-hybrid platform.Nucleic acids research · 2007Article
- SNP discovery by mismatch-targeting of Mu transposition.Nucleic acids research · 2007Article
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3 authors.
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Abstract
Bacteriophage Mu in vitro transposition constitutes a versatile tool in molecular biology, with applications ranging from engineering of single genes or proteins to modification of genome segments or entire genomes. A new strategy was devised on the basis of Mu transposition that via a few manipulation steps simultaneously generates a nested set of gene constructions encoding deletion variants of proteins. C-terminal deletions are produced using a mini-Mu transposon that carries translation stop signals close to each transposon end. Similarly, N-terminal deletions are generated using a transposon with appropriate restriction sites, which allows deletion of the 5'-distal part of the gene. As a proof of principle, we produced a set of plasmid constructions encoding both C- and N-terminally truncated variants of yeast Mso1p and mapped its Sec1p-interacting region. The most important amino acids for the interaction in Mso1p are located between residues T46 and N78, with some weaker interactions possibly within the region E79-N105. This general-purpose gene truncation strategy is highly efficient and produces, in a single reaction series, a comprehensive repertoire of gene constructions encoding protein deletion variants, valuable in many types of functional studies. Importantly, the methodology is applicable to any protein-encoding gene cloned in an appropriate vector.
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