P element screen

[T. Kaufman]

Dear Drosophilists,

As an adjunct to the Drosophila genome sequencing effort we are planning to
generate tens of thousands of P-element insertions in D. melanogaster to
obtain single P-element insertions in the vast majority of genes.   Based
on the analysis of many enhancer detector screens by the BDGP, we surmise
that a P-element can be obtained in 75% to 95% of all the Drosophila genes
(viables and lethals).   The basic strategy is to generate single P-element
insertions throughout the genome, sequence the adjacent genomic DNA, and
compare the sequence (STS) with existing genomic sequences and ESTs.  We
will keep those P-elements that are inserted in proximity of a gene: e.g.,
in the ORF, in the 5'UTR, or within a few hundred bases of the 5'UTR.
These P-element stocks will then be deposited in the Stock Center in
Bloomington Indiana.

We are planning on using simple P-elements that contain the white[+] gene.
Three different P-elements are at present being considered:

1) A P-element that contains an enhancer detector driving GAL4 (e.g., Brand
and Perrimon, 1993)

2) 2) A P-element that contains UAS sites in front of a promoter to
facilitate ectopic expression screens (e.g., Rorth, 1997)

3) 3) A P-element containing FRT sites.

We welcome your input and thoughts as we believe that this is a one time
experiment given the magnitude of the task.  If you have any suggestions,
please contact hbellen at bcm.tmc.edu before May 1, 1999. Thank You.

Hugo Bellen
Thom Kaufman
Allan Spradling
Gerry Rubin



Thom Kaufman   	     	       kaufman at sunflower.bio.indiana.edu
--- Biology Dept.,  Indiana University, Bloomington, IN 47405 ---
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