Cloning and Prokaryotic Expression of JHEH Gene from Spodoptera litura
ZHANG Lili
WU Yiqiong
YANG Zhengfei
GUO Li
WEI Jiaping
KAN Yunchao
Abstract：In order to investigate the function of juvenile hormone epoxide hydrolase(JHEH) gene in growth and development of Spodoptera litura, the open reading frame of SlJHEH was cloned by reverse transcription-polymerase chain reaction(RT-PCR) and expressed prokaryotically.The open reading frame was 1 389 bp in length, and encoded 462 amino acids.Its predicted molecular mass and isoelectric point were 52 ku and 8.68, respectively.The residues Asp226, Glu402, His429 that formed the potential catalytic triad, and the residues Tyr297, Tyr372, and the HGWP motif that constituted the potential oxyanion hole were found in the deduced amino acid sequences.Sl JHEH had a closely phylogentic relationship with JHEH of Helicoverpa armigera, with the amino acid sequence identity reaching 78％.The recombinant p ET32a-SlJHEH was constructed by ligating the code sequence of SlJHEH with prokaryotic expression vector p ET32a.Then the recombinant genetic engineering bacteria to express JHEH were obtained by transforming pET32a-Sl JHEH into competent cells of Escherichia coli BL21(DE3).The results of SDS-PAGE and Western blotting analysis showed that the expression of Sl JHEH gene in E.coli was high and accurate, which provided the promise for functional study of SlJHEH gene.
Keywords：Spodoptera lituraJuvenile hormone epoxide hydrolaseCloningSequence analysisProkaryotic expressionRecombinant protein
Publication Date：2018-12-01
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：6( 84-89 )