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Fig. 1. Schematic of strategy used to isolate Manduca slowpoke cDNA.
Degenerate oligonucleotide PCR primers designed to conserved regions of the
Slo protein were used to amplify cDNA fragments msslo A-1, A-2 and
A-3. Each of these cDNA fragments varied slightly in size due to
alternate splicing. Manduca sexta specific PCR primers were used in
conjunction with degenerate PCR primers upstream and downstream of the
A fragments to amplify cDNA fragments msslo B and msslo
C. 5' and 3' RACE was performed to attain and confirm the
ends of the cDNA sequence. When assembled, an msslo cDNA containing
alternate exons E1, G1 and I1 totals 3693 bp in length.
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