pHS22

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. [1] Genomic clone containing 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. [1] 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. [1] The hsp82 sequences begin 5 kb upstream of the transcription starting point. Expression is constitutive, but can be increased with heat. [1] Selection of transgenic Drosophila by G418 has been done with embryos, larvae, and adults. Adults can be reselected by 6% ethanol. [1] 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. [1] P-transposable (integrating) shuttle vector. [1] Restriction digests of the clone give the following sizes (kb): KpnI–16.0; XbaI–9.4, 3.1, 0.90, 0.72, 0.56, 0.30; BamHI/KpnI–7.2, 6.6, 1.3. (ATCC staff) Medium is 1227 LB plus ampicillin. Hosts: E.coli HB101, Drosophila melanogaster, E.coli.