[Drosophila] is there transposase repressor construct?
Vef, Dr. Olaf
via dros%40net.bio.net
(by ovef from uni-mainz.de)
Wed Sep 16 07:40:29 EST 2009
Dear Drosophilists
If you want to construct deletions using P-elements in trans (Adam C. Paré et al., Genetics, Vol. 181, 53-63, 2009), one have to combine one (first) p-element with the transposase source (Delta2-3). In this first step you can not avoid the mobilisation (or local hop) of this first p-element before you combine it with the second p-element.
My idea is to repress (inhibit respectively) P-element mobilization by the transposase repressor (product of the first three exons of the transposase gene).
Does anybody of you know if there is an existing transposase repressor element (containing only the first three exons of the transposase gene) inserted in genome of Drosophila? It shouldnt be able to be mobilized by the wild type P-element transposase (like the Delta2-3 element or in a piggyBac element e.g.)?
Any suggestions or critics to the idea are welcomed!
hypothetic crossing scheme:
marked chromosome first p-element / CyO ; TMS (containing Delta2-3) Sb / Lyra or Drop (marked chromosome) p-(transposase-repressor)
crossing to
marked chromosome second p-element / CyO ; +/+
collect male offspring (Sb without Lyra or Drop)
marked chromosome first p-element / marked chromosome second p-element ; TMS (containing Delta2-3) Sb /+ (in the germ line of these males the transposase induced male recombination takes place)
crossing to
Plum-balancer / CyO
collect balanced male offspring (without Sb)
-------------------marker-------------first p-element--------------------------------------------------------------------------------------------------
--------------------------------------------------------------second p-element-----------marker-----------------------------------------------------
male recombination results in
-------------------marker-------------first p-element ### second p-element-----------marker-------------------------------------------------
------------------------------------------------------balancer------------------------------------------------------------------------------------------
(deleted segment = ###)
principle of method (Adam C. Paré et al., Genetics, Vol. 181, 53-63, 2009)
Best regards,
Olaf
Dr. Olaf Vef
Institute for Genetics
Johannes Gutenberg-University, Mainz
Johann-Joachim-Becher-Weg 32
D-55128 Mainz
GERMANY
email: ovef from mail.uni-mainz.de
Tel.: 049-(0)6131-3925347
Fax: 049-(0)6131-3925845
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