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. The hsp82 sequences begin 5 kb upstream of the transcription starting point. 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. 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. 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. |
