Transposon Tn5.

Josephine Bay Paul Center for Comparative Molecular Biology and Evolution, Woods Hole, Massachusetts 02543, USA.
Annual Review of Genetics (Impact Factor: 17.44). 09/2008; 42:269-86. DOI: 10.1146/annurev.genet.42.110807.091656
Source: PubMed

ABSTRACT Tn5 was one of the first transposons to be identified ( 10 ). As a result of Tn5's early discovery and its simple macromolecular requirements for transposition, the Tn5 system has been a very productive tool for studying the molecular mechanism of DNA transposition. These studies are of broad value because they offer insights into DNA transposition in general, because DNA transposition is a useful model with which to understand other types of protein-DNA interactions such as retroviral DNA integration and the DNA cleavage events involved in immunoglobulin gene formation, and because Tn5-derived tools are useful adjuncts in genetic experimentation.

1 Bookmark
  • [Show abstract] [Hide abstract]
    ABSTRACT: The preparation of plasmid-borne RIVET libraries can be troublesome when high genomic coverages are needed. We present here the construction and functional validation of a new set of miniTn5 promoter traps to generate tnpR-based RIVET libraries. The ability to generate tnpR transcriptional fusions by transposition will significantly facilitate the setup of RIVET studies in those bacteria where Tn5 transposition is operative.
    Journal of microbiological methods 02/2013; · 2.43 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The essential molecular chaperonin GroEL is an example of a functionally highly versatile cellular machine with a wide variety of in vitro applications ranging from protein folding to drug release. Directed evolution of new functions for GroEL is considered difficult, due to its structure as a complex homomultimeric double ring and the absence of obvious molecular engineering strategies. In order to investigate the potential to establish an orthogonal GroEL system in Escherichia coli, which might serve as a basis for GroEL evolution, we first successfully individualised groEL genes by inserting different functional peptide tags into a robustly permissive site identified by transposon mutagenesis. These peptides allowed fundamental aspects of the intracellular GroEL complex stoichiometry to be studied and revealed that GroEL single-ring complexes, which assembled in the presence of several functionally equivalent but biochemically distinct monomers, each consist almost exclusively of only one type of monomer. At least in the case of GroEL, individualisation of monomers thus leads to individualisation of homomultimeric protein complexes, effectively providing the prerequisites for evolving an orthogonal intracellular GroEL folding machine.
    ChemBioChem 10/2013; · 3.74 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The history of life has been characterised by evolutionary transitions in individuality, the grouping together of independently replicating units into new larger wholes: genes to chromosomes, chromosomes in genomes, up to three genomes in cells, and cells in multicellular organisms that form groups and societies. Central to understanding these transitions is to determine what prevents selfish behaviour at lower levels from disrupting the functionality of higher levels. Here, I review work on transposable elements, a common source of disruption at the genome level, in light of the evolutionary transitions framework, and argue that the rapid influx of data on transposons from whole-genome sequencing has created a rich data source to incorporate into the study of evolutionary transitions in individuality.
    Trends in Ecology & Evolution 11/2013; · 15.39 Impact Factor