Tn5 transposition: a molecular tool for studying protein structure-function.
Transposon-based technologies are important genetic tools for global genome analysis and, as discussed in the present paper, in detailed studies of protein structure-function. Various different transposition systems can be used in these studies but this paper uses Tn5-related systems as a model. In particular, the following four different technologies are described in this paper: (i) using transposition to generate nested deletion families, (ii) using transposons to generate functional protein fusions to reporter functions, (iii) mapping protein secondary structures through the generation and analysis of in-frame linker insertions and (iv) using sequential transposition events to generate random gene fusions. The success of these forward genetic technologies requires that the transposition system be efficient and manifest near-random target sequence selection.
Department of Biochemistry, University of Wisconsin, Madison, WI 53706, USA. reznikoff@biochem.wisc.edu
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D017353:Gene Deletion; D017930:Genes, Reporter / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D019895:Transposases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D011994:Recombinant Proteins; C473503:Tn5 transposase; D019895:Transposases