Cloning, expression and functional analysis of a general odorant-binding protein 2 gene of the rice striped stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae).
A full-length cDNA encoding a general odorant binding protein 2 (GOBP2) was cloned from the antennae of the rice striped stem borer, Chilo suppressalis (Lepidoptera: Pyralidae), by the combination of reverse transcription PCR (RT-PCR) and rapid amplification of cDNA ends PCR (RACE-PCR). The cDNA contains a 489 bp open reading frame, which encodes a 162 amino acid protein, termed as Ch. suppressalis GOBP2 (CsupGOBP2). CsupGOBP2 is similar in the number of amino acids and protein sequence to GOBP2s in other species of Lepidoptera. RT-PCR results showed that CsupGOBP2 mRNA was highly expressed in the adult antennae of both females and males, as was CsupGOBP2 protein as revealed by Western blot analysis. CsupGOBP2 expressed in Escherichia coli was purified by affinity chromatography, refolding and gel filtration from the inclusion body. Fluorescence emission spectra and competitive binding assays by using N-phenyl-1-naphthylamine as first binding ligand and odorants as potential competitors revealed that the CsupGOBP2 protein has significant affinity to cis-11-hexadecenal (Z11-16:Ald), the main component of Ch. suppressalis pheromone and to laurinaldehyd and benzaldehyde, two general plant volatile aldehydes.
Gong
Z-J
ZJ
State Key Laboratory of Rice Biology, Ministry of Agriculture, Zhejiang, China.
Zhou
W-W
WW
Yu
H-Z
HZ
Mao
C-G
CG
Zhang
C-X
CX
Cheng
J A
JA
Zhu
Z-R
ZR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli; D005260:Female; D015870:Gene Expression; D019476:Insect Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008024:Ligands; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018035:Receptors, Odorant / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D019476:Insect Proteins; D008024:Ligands; D018035:Receptors, Odorant; C045648:odorant-binding protein