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The juvenile hormone binding protein of silkworm haemolymph: gene and functional analysis
Author(s) -
Vermunt A. M. W.,
Kamimura M.,
Hirai M.,
Kiuchi M.,
Shiotsuki T.
Publication year - 2001
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1046/j.1365-2583.2001.00249.x
Subject(s) - hemolymph , bombyx mori , biology , complementary dna , juvenile hormone , microbiology and biotechnology , recombinant dna , amino acid , gene , peptide sequence , biochemistry , hormone
A cDNA fragment of haemolymph juvenile hormone binding protein (hJHBP) from larvae of Bombyx mori was amplified by RT‐PCR using degenerate primers based on the N ‐terminal amino acid sequence of purified hJHBP and a conserved region near the C‐terminus of other lepidopteran hJHBPs. 5′‐ and 3′‐ends were amplified by RACE to yield cDNAs, hJHBP1 and hJHBP2, encoding 225 amino acids with three substitutions. hJHBP‐mRNA levels in the fat body were constant in the 4th instar, but decreased in the 5th. JHBP protein was constant until wandering, then declined. Recombinant hJHBP1 expressed in E. coli migrated on SDS‐PAGE with a M r of 32 kDa and showed a K d of 4.5 × 10 −7 M with JH III, both similar to those of native hJHBP.