Purification of juvenile hormone esterase and molecular cloning of the cDNA from Manduca sexta.
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 1512bp 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 2220bp, with a 1749bp coding region. The deduced protein sequence contains 573 amino acids.
Hinton
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AC
Department of Entomology and Cancer Research Center, University of California at Davis, One Shields Avenue, Davis, CA 95616, USA.
Hammock
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BD
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006458:Hemolymph / Q000201:enzymology; D018402:Manduca / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data
D018076:DNA, Complementary; D002265:Carboxylic Ester Hydrolases; C038003:juvenile hormone esterase