Purification of juvenile hormone esterase and molecular cloning of the cDNA from Manduca sexta

A. C. Hinton, B. D. Hammock

Research output: Contribution to journalArticle

30 Scopus citations

Abstract

Juvenile hormone esterase (JHE) is a highly specific enzyme important for regulating the onset of metamorphosis in lepidopteran insects. After affinity chromatography of the hemolymph proteins of Manduca sexta, the pure JHE protein was digested with Lys-C and the resultant peptides were purified by microbore HPLC. Two peptides were selected for sequencing. Based upon these amino acid sequences, degenerate RT-PCR was performed in order to amplify a partial cDNA sequence from mRNA from the fat body of M. sexta. A 1512 bp partial cDNA was generated and found to be highly homologous to the JHE from Heliothis virescens. 5′ and 3′ RACE were performed to obtain the full length cDNA sequence. The cDNA has a total length of 2220 bp, with a 1749 bp coding region. The deduced protein sequence contains 573 amino acids.

Original languageEnglish (US)
Pages (from-to)57-66
Number of pages10
JournalInsect Biochemistry and Molecular Biology
Volume32
Issue number1
DOIs
StatePublished - 2001

Keywords

  • Affinity purification
  • CDNA
  • Esterase
  • Juvenile hormone
  • Manduca sexta
  • PCR

ASJC Scopus subject areas

  • Insect Science
  • Biochemistry

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