Cloning,molecular characteristics and expression analysis of juvenile hormone binding protein gene AcinJHBP in Apocheima cinerarius
[Objective]The present paper aimed to explore the role of AcinJHBP gene in the growth,development and diapause of Apocheima cinerarius.[Method]Based on the transcriptome data(non-diapause pupa/diapause pupa and diapause pupa/diapause depupa),JHBP gene was cloned and named as AcinJHBP(GenBank entry number:OR860357).The sequence and physicochemical properties of AcinJHBP gene were analyzed by bioinformatics software.The temporal and spatial expression of AcinJHBP(different developmental stages and tissues)was detected by qPCR.[Result]The complete CDS sequence of AcinJHBP gene of A.cinerarius was 726 bp,encoding 241 amino acids,the pre-dicted molecular weight of protein was 26.97 kDa,and the predicted isoelectric point was 5.13.The predicted molecular formula of AcinJH-BP gene was C1211 H1915 N313 O364S9,the total charge of negative charge residues(Asp+Glu)was 34,the total charge of positive charge res-idues(Arg+Lys)was 26,the half-life and aliphatic index were 30 hours and 97.51,respectively.The average hydrophilicity of the protein was-0.098,and the predicted instability coefficient was 27.13.Therefore,the protein was a hydrophilic stable protein,and the N-termi-nal amino acid was methionine(Met,M).AcinJHBP contained a signal peptide of 17 amino acid residues at the N-terminus,but had no transmembrane region.Phosphoric acid sites were found in serine,threonine and tyrosine phosphate sites were 9,6 and 3,respectively,and no glycosylation sites were found.Subcellular localization results showed that it was mainly located outside the cell.Phylogenetic analysis showed that AcinJHBP had the closest relationship with the hemolymph juvenile hormone binding protein JHBP of Operophtera brumata.The results of gene expression showed that the expression of AcinJHBP was lower at different developmental stages(except the 3 rd instar larvae),and there was no significant difference between the egg and the 5th instar larvae(P＞0.05),for all that the expression of AcinJHBP showed a decreasing trend in non-diapause pupa-adult stage.In different tissues,the expression of AcinJHBP gene was the highest in the male head of A.cinerarius.[Conclusion]AcinJHBP gene is closely related to the growth,development and diapause of A.cinerarius.The results lay a theoretical foundation for further revealing the function of the gene in the summer and winter diapause of the pupae in A.cinerarius.
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