Cloning and spatio-temporal expression of the odorant binding protein ComOBP1 gene from Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae)
The odorant binding protein gene CmonOBP1 (Genbank number:KU170686) from Cryptolaemus montrouzieri Mulsant was cloned by technology of RT-PCR and RACE.The full length was 922 bp,containing a 5' UTR (untranslated region) of 40 bp and a 3' UTR of 462 bp.ORF (Open Reading Frame) was 420 bp which encoded a polypeptide of 139 amino acids with an estimated molecular weight of 15.516 kDa and pI of 6.57.Besides,the sequence had an AATAAA tailing signal.The predicted N-terminal hydrophobic region containing 20 amino residues displayed the characteristic features of a signal peptide,and no transmembrane domain was predicted.Four conservative cysteine residues classified CmonOBP1 as the first Minus-C OBP which was found in C.montrouzieri.The amino acid sequence had only one N-glycosylation site 62 NLSA,and two potential phosphorylation sites.Compared with other insect Minus-C OBPs,CmonOPB1 was much identitier to the other Coleoptera.By Real-time PCR and RT-PCR methods,we determined the expression pattern of CmonOBP1 in different developmental stages,various adult tissues,different larval food regimes and other food.Spatio-temporal expression result showed that CmonOBP1 expressed in every stage and the male adult had the highest level.In addition,CmonOPB1 had a higher expression level in adult head and wing than that of other adult tissues.When larval diet changed either poor or rich,the expression level wouldn't be influenced markedly,while when the food changed from natural prey Planococcus citri Risso to Megoura japonica Matsumura,the expression level decreased significantly.The result suggested that the expression level of CmonOBP1 gene was development,tissues and prey type dependent,indicating a distinct sensitivity in olfactory.In addition,CmonOBP1 gene might play an important role in the process of male adult feeling related pheromone.
NETL
NSTL
万方数据
维普
