1. P Element Transposition In Vitro Proceeds by a Cut-and-Paste Mechanism and Uses GTP as a Cofactor David Coates

2. Introduction

3. Introduction
Elucidate the mechanism of P Element transposition in vitro

4. Introduction
Elucidate the mechanism of P Element transposition in vitro
Examine the effects of adding possible transposition cofactors (besides transposase)

5. Introduction
Elucidate the mechanism of P Element transposition in vitro
Examine the effects of adding possible transposition cofactors (besides transposase)
Find optimal mechanism

6. Introduction
Elucidate the mechanism of P Element transposition in vitro
Examine the effects of adding possible transposition cofactors (besides transposase)
Find optimal mechanism
Possible result of being able to insert sequences in vitro

7. Genetic Assay for P Element Transposition In Vitro
Overview: Recombinant P Element constructed with tetr gene between P Element Terminal sequences, designated P1-Pwt-tet.
These were used because of their complete lack of homology with the target plasmid.
Transposase was bound, then target DNA added, and the results of the reaction were electroporated into E. coli on ampicillin agar and ampicillin/tetracycline agar. Ratio of colonies amprtetr/colonies ampr taken.

9. Genetic Assay for P Element Transposition In Vitro