Structure And Function Analysis Of General Odor Binding Protein In Eogystia Hippophaecolus

Eogystia hippophaecolus belongs to the Eogystia genus of Cossidae,Lepidoptera,which distributes in Inner Mongolia and Liaoning.The main stem and root of Hippophae rhamnoides Linn.are damaged by larvae,which seriously damages the ecological and economic benefits of the seabuckthorn forest.Odorant binding proteins(OBPs)are the first key proteins in insect signal transduction,which play an important role in selective recognition and transportation of odor molecules.In this study,we measured the expression patterns of 13 OBPs genes in antennae,lower lip,foot(anterior,middle and posterior)and genitalia of male and their sequence characteristics.Prokaryotic expression and western blotting assays were used to detect the expression characteristics of the protein and verify the correct expression of the obtained protein.By analyzing the expression of EhipGOBPs gene and protein levels comprehensively,the function of EhipGOBPs was revealed from both gene and protein levels.This study provides more scientific basis and technical support for the further application of host plant volatiles and sex pheromones to coordinate the prevention and control of the harm of E.hippophaecolus moth.The results are as follows:1.The expression patterns of 13 OBPs genes in antennae,feet(anterior,middle and posterior),lower lip whiskers and external genitals of male adults were clarified,and the study on the expression patterns of OBPs in different sensory tissues of E.hippophaecolus was further improved.Thirteen OBPs were expressed in four tissues,and the expression levels were different.Antennae,feet and genitals are the main parts of OBPs expression.Eight OBPs expressed more in antennae than the other three tissues,while OBP12,OBP16,OBP17 and OBP24 expressed relatively high in feet;OBP19 expressed specifically in external genitals;OBPs expressed relatively low in lower lip whiskers,and no biased genes were found.2.Two complete open reading frame bases and amino acid sequences of EhipGOBPs gene were obtained.The three-dimensional structures of EhipGOBP1 and EhipGOBP2 proteins were simulated.The complete cNDA of EhipGOBP1 gene is 600bp in size,encoding 199 amino acids,and its molecular weight is predicted to be 23KDa.The complete cNDA length of EhipGOBP2 gene is 501bp,encoding 166 amino acids,and the molecular weight of protein is 18.9KDa.Amino acid sequence analysis showed that the similarity between EhipGOBP1 and EhipGOBP2 was 41.2%.EhipGOBPs contain 7 a helices,and the three pairs of disulfide bonds formed by six cysteines support their structure.3.The expression of EhipGOBPs protein in vitro was successfully realized,and the expression patterns of EhipGOBPs protein in antennae,feet(anterior,middle and posterior),lower lip whiskers and external genitals of male adults were clarified.The target fragment was confirmed by restriction enzyme digestion.The recombinant EhipGOBPs protein was expressed in inclusion bodies.The size of the band was consistent with the target fragment by gel electrophoresis,indicating that the EhipGOBPs protein was expressed in vitro.The polyclonal antibodies against EhipGOBPs were used for Western blotting of four different tissues of male adults of E.hippophaecolus.The results showed that EhipGOBPs protein was expressed in all four tissues.