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. 1989 Oct;86(20):8123–8127. doi: 10.1073/pnas.86.20.8123

Isolation and expression of the eclosion hormone gene from the tobacco hornworm, Manduca sexta.

F M Horodyski 1, L M Riddiford 1, J W Truman 1
PMCID: PMC298227  PMID: 2813382

Abstract

Eclosion hormone (EH) is a 62-amino acid neuropeptide that initiates the ecdysis behavior of insects. The EH-encoding gene of the tobacco hornworm, Manduca sexta, was isolated by using a designed 72-mer oligonucleotide probe. Sequence analysis of this gene and its corresponding cDNA showed that the EH gene is 7.8 kilobases and consists of three exons. Exon I is totally nontranslated; exon II contains a 26-amino acid signal peptide and amino acids 1-4 of the EH peptide, and exon III encodes the remainder of the peptide. The EH gene is present in a single copy per haploid genome and transcribes an 0.8-kb mRNA that is expressed in larval, diapausing pupal, and developing adult brains but not in the ventral nerve cord or in nonneural tissues. In situ hybridization showed that the EH gene is expressed in two pairs of ventromedial neurosecretory cells in brains of both larvae and developing adults.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson S., Kingston I. B. Isolation of a genomic clone for bovine pancreatic trypsin inhibitor by using a unique-sequence synthetic DNA probe. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6838–6842. doi: 10.1073/pnas.80.22.6838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  4. Copenhaver P. F., Truman J. W. Identification of the cerebral neurosecretory cells that contain eclosion hormone in the moth Manduca sexta. J Neurosci. 1986 Jun;6(6):1738–1747. doi: 10.1523/JNEUROSCI.06-06-01738.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dale R. M., McClure B. A., Houchins J. P. A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18 S rDNA. Plasmid. 1985 Jan;13(1):31–40. doi: 10.1016/0147-619x(85)90053-8. [DOI] [PubMed] [Google Scholar]
  6. Douglass J., Civelli O., Herbert E. Polyprotein gene expression: generation of diversity of neuroendocrine peptides. Annu Rev Biochem. 1984;53:665–715. doi: 10.1146/annurev.bi.53.070184.003313. [DOI] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  8. Grantham R., Gautier C., Gouy M., Jacobzone M., Mercier R. Codon catalog usage is a genome strategy modulated for gene expressivity. Nucleic Acids Res. 1981 Jan 10;9(1):r43–r74. doi: 10.1093/nar/9.1.213-b. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  10. Gubler U. Second-strand cDNA synthesis: mRNA fragments as primers. Methods Enzymol. 1987;152:330–335. doi: 10.1016/0076-6879(87)52038-9. [DOI] [PubMed] [Google Scholar]
  11. Horodyski F. M., Riddiford L. M. Expression and hormonal control of a new larval cuticular multigene family at the onset of metamorphosis of the tobacco hornworm. Dev Biol. 1989 Apr;132(2):292–303. doi: 10.1016/0012-1606(89)90226-1. [DOI] [PubMed] [Google Scholar]
  12. Hultmark D., Klemenz R., Gehring W. J. Translational and transcriptional control elements in the untranslated leader of the heat-shock gene hsp22. Cell. 1986 Feb 14;44(3):429–438. doi: 10.1016/0092-8674(86)90464-2. [DOI] [PubMed] [Google Scholar]
  13. Kataoka H., Troetschler R. G., Kramer S. J., Cesarin B. J., Schooley D. A. Isolation and primary structure of the eclosion hormone of the tobacco hornworm, Manduca sexta. Biochem Biophys Res Commun. 1987 Jul 31;146(2):746–750. doi: 10.1016/0006-291x(87)90592-4. [DOI] [PubMed] [Google Scholar]
  14. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  15. Marti T., Takio K., Walsh K. A., Terzi G., Truman J. W. Microanalysis of the amino acid sequence of the eclosion hormone from the tobacco hornworm Manduca sexta. FEBS Lett. 1987 Jul 27;219(2):415–418. doi: 10.1016/0014-5793(87)80263-6. [DOI] [PubMed] [Google Scholar]
  16. Maruyama T., Gojobori T., Aota S., Ikemura T. Codon usage tabulated from the GenBank genetic sequence data. Nucleic Acids Res. 1986;14 (Suppl):r151–r197. doi: 10.1093/nar/14.suppl.r151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nambu J. R., Murphy-Erdosh C., Andrews P. C., Feistner G. J., Scheller R. H. Isolation and characterization of a Drosophila neuropeptide gene. Neuron. 1988 Mar;1(1):55–61. doi: 10.1016/0896-6273(88)90209-7. [DOI] [PubMed] [Google Scholar]
  19. O'Shea M., Schaffer M. Neuropeptide function: the invertebrate contribution. Annu Rev Neurosci. 1985;8:171–198. doi: 10.1146/annurev.ne.08.030185.001131. [DOI] [PubMed] [Google Scholar]
  20. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  21. Pustell J., Kafatos F. C. A convenient and adaptable package of computer programs for DNA and protein sequence management, analysis and homology determination. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):643–655. doi: 10.1093/nar/12.1part2.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Riddiford L. M. Changes in translatable mRNAs during the larval-pupal transformation of the epidermis of the tobacco hornworm. Dev Biol. 1982 Aug;92(2):330–342. doi: 10.1016/0012-1606(82)90179-8. [DOI] [PubMed] [Google Scholar]
  23. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sarkar G., Sommer S. S. Access to a messenger RNA sequence or its protein product is not limited by tissue or species specificity. Science. 1989 Apr 21;244(4902):331–334. doi: 10.1126/science.2565599. [DOI] [PubMed] [Google Scholar]
  26. Scheller R. H., Jackson J. F., McAllister L. B., Rothman B. S., Mayeri E., Axel R. A single gene encodes multiple neuropeptides mediating a stereotyped behavior. Cell. 1983 Jan;32(1):7–22. doi: 10.1016/0092-8674(83)90492-0. [DOI] [PubMed] [Google Scholar]
  27. Schneider L. E., Taghert P. H. Isolation and characterization of a Drosophila gene that encodes multiple neuropeptides related to Phe-Met-Arg-Phe-NH2 (FMRFamide). Proc Natl Acad Sci U S A. 1988 Mar;85(6):1993–1997. doi: 10.1073/pnas.85.6.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schulz-Aellen M. F., Roulet E., Fischer-Lougheed J., O'Shea M. Synthesis of a homodimer neurohormone precursor of locust adipokinetic hormone studied by in vitro translation and cDNA cloning. Neuron. 1989 Apr;2(4):1369–1373. doi: 10.1016/0896-6273(89)90075-5. [DOI] [PubMed] [Google Scholar]
  29. Schwartz L. M., Truman J. W. Hormonal control of rates of metamorphic development in the tobacco hornworm Manduca sexta. Dev Biol. 1983 Sep;99(1):103–114. doi: 10.1016/0012-1606(83)90257-9. [DOI] [PubMed] [Google Scholar]
  30. Sossin W. S., Fisher J. M., Scheller R. H. Cellular and molecular biology of neuropeptide processing and packaging. Neuron. 1989 May;2(5):1407–1417. doi: 10.1016/0896-6273(89)90186-4. [DOI] [PubMed] [Google Scholar]
  31. Teyke T., Weiss K. R., Kupfermann I. A subpopulation of cerebral B cluster neurones of Aplysia californica is involved in defensive head withdrawal but not appetitive head movements. J Exp Biol. 1989 Nov;147:1–20. doi: 10.1242/jeb.147.1.1b. [DOI] [PubMed] [Google Scholar]
  32. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  33. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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