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Yashu Zhang,Xiao Zhao,Jinyun Ji,Tinghao Kang,Jian Hong Li,Hu Wan 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.2
In the present study, a phenylalanine hydroxylase (PAH) was cloned and characterized from the common cutworm Spodoptera litura (SlPAH). The SlPAH gene is an 1863 bp full-length cDNAwith a 1371 bp open reading frame that encodes a predicted 52.35 kDa protein of 457 amino acid residueswith an isoelectric point of 5.39. The SlPAH gene contains 6 exons and 5 introns. A multiple sequence analysis of SlPAH showed an 88–68% sequence identity to Papilio xuthus PAH and other insect PAHs. A phylogenetic tree revealed that SlPAH is a novel member of the PAH family. Tissue distribution analyses indicated that SlPAH is constitutively expressed in the epidermis, midgut, and fat body, with the highest expression occurring in the fat body on days 2 and 3 of the sixth-instar larval stage and day 1 of the prepupa stage. However, RNAi-mediated SlPAH gene silencing had no observable phenotype in S. litura larvae. Additionally, SlPAH expression is acutely up-regulated in the fat body after injection with Bacillus thuringiensis, Escherichia coli, or Beauveria bassiana. Therefore, these results provide a molecular basis for the functional roles of SlPAH during development and the innate immune response in S. litura.
Hu Wan,Yashu Zhang,Xiao Zhao,Hong You,Jian Hong Li,Jinyun Ji 한국응용곤충학회 2015 Journal of Asia-Pacific Entomology Vol.18 No.3
Insect cytokine growth-blocking peptides (GBPs) play functional roles in growth regulation and the innate immune response.Moreover, high concentrations of GBPs significantly induce larvalmorphological abnormalities and lethality. In the present study, a recombinant AcNPV, AcNPV-SeGBPm, expressing a growth-blocking peptide fromthe beet armywormSpodoptera exigua (SeGBP) under the control of a polyhedrin promoterwas constructed. Western blot analysis indicated that the mature peptide of SeGBP (SeGBPm) is expressed as a 3-kDa peptide in recombinant baculovirus-infected Sf9 insect cells. SeGBPm expression in Sf9 insect cells decreased cell viability significantly and induced apoptotic cell death. In addition, the body weights of S. exigua larvae inoculated with AcNPV-SeGBPm decreased significantly compared to those of larvae inoculated with wild-type AcNPV. Moreover, AcNPV-SeGBPm improved the rate of larvae killing (LT50) by approximately 1 day compared to wild-type AcNPV. Taken together, the obtained data demonstrate the great potential for future investigations of growth-blocking peptides for use as biopesticides.