Cloning and differential expression of the Apis cerana cerana and Apis mellifera ligustica odorant binding protein gene OBP12
Author of the article:DU Ya-Li1** FENG Yu-Jia1** MA Wei-Hua2 TAI Miao-Miao1 LI Xin-Yu1 SU Wen-Ting1 ZHAO Hui-Ting3 J
Author's Workplace：(1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China; 2. Institute of Horticulture, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China; 3. College of Life Science, Shanxi Agricultural University, Taigu 030801, China)
Key Words：Apis cerana cerana; Apis mellifera ligustica; OBP12; gene cloning; bioinformatics analysis; temporal-spatial expression
Abstract：
[Objectives] To clone the cDNA sequence of OBP12 from Apis cerana cerana and Apis mellifera ligustica, predict their protein structures and compare
differences in gene expression in different tissues and developmental stages
between these two species. [Methods] The full-length cDNA sequence of OBP12 was amplified and cloned from
antennae of A. c. cerana and A. m. ligustica using RT-PCR.
Its physiochemical properties and structural characteristics are described and
a phylogenetic tree of the deduced amino acids constructed using bioinformatics
software. The expression profiles of AcerOBP12
and AmelOBP12 mRNA in different
tissues (antenna, head, thorax, abdomen, legs and wings) at different
developmental stages (1, 5, 10, 15, 20, 25 and 30 d) were detected with
real-time PCR and compared. [Results] The entire ORF sequence of AcerOBP12 and AmelOBP12 containing 453 bp was successfully obtained. Both genes
encoded a putative protein of 150 amino acids with estimated molecular weights
of 17.76 ku and 17.42 ku, respectively. Both deduced proteins had a signal
peptide of 22 amino acids at the N-terminal region, no transmembrane structure
and contained six conserved cysteine sites, suggesting that they belong to the
Classical OBP subfamily. Both genes had 6 α-helices linked by three disulfide
bonds. The phylogenetic tree indicates that AcerOBP12 and AmelOBP12 belong to
the same group, clustering together with the Hymenopteran genes HlabPBPGp-9-like,
HsalPBPGp-9-like, LhumPBPGp-9-like and WaurPBPGp-9-like. Real-time PCR revealed
that OBP12 was expressed in various
tissues at different developmental stages of A. c. cerana and A. m.
ligustica, and that expression profiles in
antennae were significantly higher than in other tissues (P<0.01). In other tissues, OBP12
was primarily expressed in the legs and wings and only weakly expressed in the
head, thorax and abdomen. Expression
levels of OBP12 in different
tissues and developmental stages of A. m. ligustica were higher than in the corresponding tissues and developmental
stages of A.
c. cerana. [Conclusion] AcerOBP12 and AmelOBP12 belong to the
classical OBP subfamily and may be pheromone binding proteins (PBPs). Tissue
expression profiles suggest that OBP12
may have a gustatory function,
and play a role in olfaction.