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Introduction

P Element Transposition In Vitro Proceeds by a Cut-and-Paste Mechanism and Uses GTP as a Cofactor David Coates. Introduction. Introduction. Elucidate the mechanism of P Element transposition in vitro. Introduction. Elucidate the mechanism of P Element transposition in vitro

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Introduction

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  1. P Element Transposition In Vitro Proceeds by a Cut-and-Paste Mechanism and Uses GTP as a CofactorDavid 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 tetrgene 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.

  8. Genetic Assay for P Element Transposition In Vitro

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