Molecular cloning and bacterial expression of pheromone binding protein in the antennae of Helicoverpa armigera (Hübner).
A cDNA clone coding for pheromone binding protein was isolated from the antennae of Helicoverpa armigera by RT-PCR and (5'/3')-RACE technique. The full-length of H. armigera pheromone binding protein (HarmPBP) was 952 bp, possessing 162 amino acid residues including a signal peptide of 20 amino acids. Its predicted molecular weight and isoelectric point were 18.26 kDa and 5.23, respectively. This deduced amino acid sequence shared some common structural features with odorant-binding proteins from several moth species, including the six conserved cysteine motif, a typical characteristic of insect's odorant-binding proteins. Northern blot showed that HarmPBP is specifically expressed in the antennae of Helicoverpa armigera and more abundantly expressed in male than female. During the antennal development, HarmPBP is first expressed about 4 days prior to adult eclosion and rises to a plateau 2 days prior to adult eclosion. In order to obtain sufficient PBP for further determining its biochemical and physiological properties, a bacterical expression vector of PBP was constructed and successfully expressed in Escherichia coli. The recombinant PBP was shown to cross-react with an anti-PBP antiserum from Antheraea polyphemus. Polyclonal antibodies against HarmPBP were used to mark the distribution of the protein in olfactory sensilla. Very strong labeling was observed in the sensillum lymph of the hair lumen and of the sensillum-lymph cavity. In the male, HarmPBP is expressed in sensilla trichodea and not in sensilla basiconica, while in the female, it is expressed both in sensilla basiconica and sensilla trichodea.
Wang
Gui Rong
GR
State Key Laboratory of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, PR China.
Wu
Kong Ming
KM
Guo
Yu Yuan
YY
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D017344:Genes, Insect / Q000235:genetics*; D005822:Genetic Vectors; D007150:Immunohistochemistry; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D012313:RNA / Q000302:isolation & purification; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012679:Sense Organs / Q000737:chemistry / Q000502:physiology*; D016415:Sequence Alignment
D002352:Carrier Proteins; D018076:DNA, Complementary; D019476:Insect Proteins; D011994:Recombinant Proteins; C403713:pheromone binding protein, insect; D012313:RNA