Tn5 as a molecular genetics tool: In vitro transposition and the coupling of in vitro technologies with in vivo transposition.
The development of in vitro transposition technologies have provided many powerful tools for the molecular genetics research laboratory. In this chapter we describe some of these tools with a focus on the Tn5 transposition system. Tn5 technologies are particularly useful because the Tn5 transposition system has simple requirements, is efficient, random in target recognition, and robust. In particular we will describe the use of in vitro Tn5 transposition in transposon tagging and in the generation of nested deletions. We will also describe a unique in vitro/in vivo technology in which Tn5 inserts can be generated in a wide spectrum of bacterial species through the electroporation of preformed tranposase-transposon DNA complexes.
Reznikoff
William S
WS
Department of Biochemistry, University of Wisconsin, Madison, USA.
Goryshin
Igor Y
IY
Jendrisak
Jerry J
JJ
D001483:Base Sequence; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D018274:Electroporation / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D016680:Genome, Bacterial; D066298:In Vitro Techniques; D008967:Molecular Biology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011995:Recombination, Genetic / Q000235:genetics; D019895:Transposases / Q000235:genetics* / Q000378:metabolism*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D019895:Transposases