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. Expression is constitutive, but can be increased with heat. Selection of transgenic Drosophila by G418 has been done with embryos, larvae, and adults. Adults can be reselected by 6% ethanol. 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. P-transposable (integrating) shuttle vector. 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. 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. 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. Differs from pHS22 (ATCC 37840) by the removal of hsp82 upstream sequences from approximately -5000 to -868 (where the transcription start point is +1). |
