Constructing UAS-tTA; TRE70W1 or 2 We want to combine the UAS-tTA from Bruno Bello with our TRE70W.  Bruno provided the UAS-tTA in flies that also contain tetO-lacZ, so we essentially need to replace tetO-lacZ with our TRE70W1 or 2.  The following scheme should accomplish this.  Note that I have only designated the desire progeny in each generation so be certain to select the correct phenotype.

¡¡ yw;UAS-tTA/UAS-tTA;tetO-lacZ/tetO-lacZ  x  ¢¢ w;Sp/SM6a;Gl/TM3 n ¡¡ yw/w;UAS-tTA/SM6a;tetO-lacZ/TM3  x  ¢¢ w;Sp/SM6a;Gl/TM3 m w;UAS-tTA/SM6a;Gl/TM3

¡¡ w; +/+;TRE70W/TRE70W x  ¢¢ w;Sp/SM6a;Gl/TM3  n ¡¡ w; +/SM6a;TRE70W/TM3 x  ¢¢ w;Sp/SM6a;Gl/TM3 m w; Sp/SM6a;TRE 70W /TM3

¡¡ w; UAS-tTA/SM6a; Gl/TM3 x ¢¢ w; Sp/SM6a; TRE 70W /TM3 m ¡¡ w; UAS-tTA/SM6a; TRE 70W /TM3 x ¢¢ w; UAS-tTA/SM6a; TRE 70W /TM3 m w; UAS-tTA/UAS-tTA; TRE 70W /TRE 70W

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Testing for tTA induction of TRE70W Crossing UAS-tTA/UAS-tTA; TRE 70W /TRE 70W with Mz-1087.hx should result in expression of tTA in salivary glands.  In turn, this should activate expression of TRE70W.  If this is observed, we are in a position to determine if the presence of doxycycline will inhibit induction.  A simple test can be done by raising the larvae on doxycycline and determining if this represses TRE70W expression.