pHS24

Differs from pHS22 (ATCC 37840) by the removal of hsp82 upstream sequences from approximately -5000 to -868 (where the transcription start point is +1). Also contains a complete alcohol dehydrogenase coding region from D. melanogaster (3.2 kb XbaI fragment) which includes 0.66 kb of 5′ and 0.64 kb of 3′ DNA, and has all the cis-acting sequences necessary for expression. Contains hsp82-neo fusion gene which encodes the first 20 aa of D. pseudoobscura hsp82, 5 codons in a linker, fused to the fifth codon of bacterial neomycin phosphotransferase from Tn5, and followed by polyadenylation signals from D. melanogaster hsp82. The order of the major components of this construct are:3′ P-element, 5′-Adh-3′, 5′-hsp82-neo fusion-hsp82 polyadenylation signal-3′, KpnI site, and 5′ P-element. Expression is constitutive, but can be increased with heat. Can be used in Drosophila spp. cell culture transfection assays, and may function in other animal and plant systems because the hsp82 promoter is highly conserved. Selection of transgenic Drosophila by G418 has been done with embryos, larvae, and adults. Adults can be reselected by 6% ethanol. P-transposable (integrating) shuttle vector. Restriction digests of the clone give the following sizes (kb): KpnI–12.5; XbaI–11.0, 0.72, 0.50, 0.37; BamHI/KpnI–7.6, 3.8, 1.3. (ATCC staff) Medium is 1227 LB plus ampicillin. Hosts: E.coli HB101, Drosophila melanogaster, E.coli.