Cloning, bioinformatics and expression of the juvenile hormone epoxide hydrolase gene in Aspongopus chinensis Dallas
Author of the article:ZHOU Wen-Zhen WU You-Fang GUO Jian-Jun
Author's Workplace:Institute of Entomology, Guizhou University, Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region, Guiyang 550025, China
Key Words:juvenile hormone epoxide hydrolase; Aspongopus chinensis; gene clone; bioinformatics analysis; expression analysis
Abstract:
[Objectives] To sequence the Aspongopus chinensis juvenile hormone epoxide hydrolase and use
bioinformatic analysis to investigate its role in the development of A. chinensis. [Methods] The cDNA sequence of AcJHEH was amplified and cloned from adult A.
chinensis with RT-PCR. Its physiochemical properties and structural
characteristics were described and a phylogenetic tree of the deduced amino
acids constructed using bioinformatics software. The expression of AcJHEH in different tissues (ovary, testis, fat, muscle, body wall, midgut and
head) and in different developmental stages (1st, 2nd, 3rd, 4th, 5th instar larvae, adult males and females,
diapausing and reproductive adults) was measured using quantitative real-time
PCR. [Results] The entire ORF sequence of AcJHEH containing 1 363 bp was obtained. Both genes encoded a putative protein of 453
amino acids with an estimated molecular weight of 51.74 ku. The isoelectric
point (pI) was 7.66 and the molecular protein formula was C2414H3683N581O653S14.
The amino acid encoded by JHEH contained the N-terminal transmembrane motif XWG, a catalytic triad, the
conserved motif HGWP and two tyrosines, suggesting that it belongs to the epoxide hydrolase family. The phylogenetic tree indicates that AcJHEH is most similar to Halyomorpha halys JHEH and that it clusters with JHEH genes from the Hemipteran species Cimex lectulariu and Apolygus lucorum. Quantitative real-time
PCR revealed that AcJHEH is expressed in various tissues
and developmental stages of A. chinensis. Expression in the
fat body was significantly higher than in other tissues (P<0.001). AcJHEH was
most highly expressed in diapausing A. chinensis, followed by 4-5
instar larvae, in which expression was significantly higher
than in other instars (P<0.001). [Conclusion] The JHEH of A. chinensis belongs to the epoxide hydrolase enzyme family
and was identified as a juvenile hormone epoxide hydrolase. qPCR results
suggest it is involved in the molting and development of 4-5th instar larvae, the development of the adult reproductive system and diapause.