Abstract
A gene sequence encoding the neuropeptide adipokinetic hormone (AKH) was isolated from a genomic library of the tobacco hornworm, Manduca sexta. 2.5 x 10(5) recombinant plaques were screened with a 29-mer oligonucleotide substituted with deoxyinosine at six ambiguous codon positions. One clone was obtained from which we deduced an intronless gene encoding the AKH precursor (prepro-AKH). From NH2 to COOH terminus, prepro-AKH consists of a signal peptide, a single AKH peptide block followed by a Gly-Lys-Arg processing site, and a 34-residue sequence that appears unrelated to known peptides. The cloned AKH gene sequence hybridized to one transcript from the adult corpus cardiacum/corpus allatum complex. The Manduca genome appears to contain one copy of the AKH gene. These studies are a basis for explaining synthesis of a representative from a widespread, physiologically important family of arthropod neuropeptides.
Highlights
Adipokinetic Hormone Gene Sequence fromManduca sexta*One clone was obtained from which we deduced an intronless gene encoding the AKH precursor (prepro-AKH).From NH2 to COOH terminus, prepro-AKH consists of a signal peptide, a single AKH peptide block followed by a Gly-Lys-Arg processing site, and a 34-residue sequence that appears the sequence complexity of the genomic library, the short amino acid sequence (nonapeptide) available for hybridization probe design, and a high level of codon degeneracy,we isolated a single-copy AKH gene sequence using a deoxyinosine-substituted [4]oligonucleotide
A gene sequence encoding the neuropeptide adipo- amount of neuroendocrine tissue was available, and we did kinetic hormone (AKH) was isolated from a genomic not know when AKH gene transcripts were present
One clone was obtained from which we deduced an intronless gene encoding the AKH precursor.From NH2 to COOH terminus, prepro-AKH consists of a signal peptide, a single AKH peptide block followed by a Gly-Lys-Arg processing site, and a 34-residue sequence that appears the sequence complexity of the genomic library, the short amino acid sequence available for hybridization probe design, and a high level of codon degeneracy,we isolated a single-copy AKH gene sequence using a deoxyinosine-substituted [4]oligonucleotide
Summary
One clone was obtained from which we deduced an intronless gene encoding the AKH precursor (prepro-AKH).From NH2 to COOH terminus, prepro-AKH consists of a signal peptide, a single AKH peptide block followed by a Gly-Lys-Arg processing site, and a 34-residue sequence that appears the sequence complexity of the genomic library, the short amino acid sequence (nonapeptide) available for hybridization probe design, and a high level of codon degeneracy,we isolated a single-copy AKH gene sequence using a deoxyinosine-substituted [4]oligonucleotide. The Man- Oligonucleotide Probe Design-The structure of Manduca AKH is duca genome appears to contain one copy of the AKH
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