Identification And Expression Of Odorant-Binding Protein Genes From Chinese White Pine Beetle(Dendroctonus Armandi)

Olfactory systems play crucial roles in the survival and reproductive success of insects,which remarkably sensitive and selective sense of smell allows the insects to detect extremely low amounts of relevant odorants and thereby lead to various behavioral responses,e.g.,looking for food and habitat,mating,laying of eggs,avoiding predators and so on.Recognizing and binding to the odorants is the first step of insect olfaction response,in which,odorant binging proteins play key roles.Until today,there has no report about OBP gene of Chinese white pine beetle(Dendroctonus armandi),there for it is very important to research OBPs of D.armandi.In this paper we studied on the OBP gene in the antenna of D.armandi.The main results are as follows:1.We had cloned and sequensed a β-actin gene of D.armandi.A phylogenetic tree based on the amino acid sequences was constructed.The results of real-time quantitative PCR indicated that it expressed stability in the different ages and in the different oranges of adult D.armandi.The conclusion is that the β-actin gene of D.armandi can be used as a housekeeping gene.2.We cloned and sequenced an OBP gene from D.armandi.Sequence analysis showed that it belongs to PBP-GOBP family and named as DarmOBP1.Phylogenetic tree of DarmOBP1 based on predicted amino acid sequence was constructed.The result of protein structure predicted shows DarmOBP1 gene can encoding a protein with 13.54 KD.3.We constructed a prokaryotic expression vector.The vector can express a protein with 14 KD under induction of IPTG which is accordant to the predicted results.Inducing conditions are as follows,Induction temperature,28℃;IPTG concentration,1.0 mmol/L;Induction time,6 h.