Using a differential display of mRNA technique we discovered that the juvenile hormone (JH) esterase gene (Cfjhe) from Choristoneura fumiferana is directly induced by juvenile hormone I (JH I), and the JH I induction is suppressed by 20-hydroxyecdysone (20E). To study the mechanism of action of these two hormones in the regulation of expression of this gene, we cloned the 1270-bp promoter region of the Cfjhe gene and identified a 30-bp region that is located between ؊604 and ؊574 and is sufficient to support both JH I induction and 20E suppression. This 30-bp region contains two conserved hormone response element half-sites separated by a 4-nucleotide spacer similar to the direct repeat 4 element and is designated as a putative juvenile hormone response element (JHRE). In CF-203 cells, a luciferase reporter placed under the control of JHRE and a mini- (6), juvenile hormone esterase (7,8), calmodulin (9), vitellogenin (10), and several others have been reported (11,12). Through indirect action, JH was shown to modulate 20-hydroxyecdysone (20E) action by affecting the expression of genes in a 20E-induced cascade (13,14). Totally different from the above genomic actions, in ovarian follicular epithelium, JH acts through a membrane receptor to bring about rapid enzyme activation without the need for new transcription (15).Numerous attempts have been made to identify JH receptors. Palli et al. (16) used human retinoic acid receptor cDNA as a probe and identified a steroid/thyroid superfamily member from Manduca sexta. Further characterization of this cDNA revealed that this is not a JH receptor but, rather, an ecdysoneinduced transcription factor that plays a critical role in ecdysone signal transduction and is related to Drosophila hormone receptor 3 (17), subsequently named Manduca hormone receptor 3 (18). The 29-kDa nuclear protein identified in M. sexta epidermis turned out to be a low affinity JH-binding protein (19,20).The mammalian retinoid X receptor (RXR) forms a heterodimer with several nuclear receptors including the farnesoid X-activated receptor (FXR). JH III but not JH acid or methoprene can bind/activate the FXR and RXR heterodimer (21). Methoprene and methoprene acid but not JH III can activate RXR (22). These two studies suggested that RXR or its insect homologue ultraspiracle (USP) could play an important role in signal transduction of JH or JH-related compounds. Jones and Sharp (23) showed that both JH III and JHB 3 bind to a USP homodimer from Drosophila melanogaster. Subsequent studies showed that USP from D. melanogaster can bind to the DR12 response element and a reporter gene placed under the control of the DR12 response element fused to the jhe core promoter was induced by JH III (24).A D. melanogaster mutant tolerant to methoprene (Met) was identified (25). An 85-kDa protein isolated from Met flies showed a 6-fold lower affinity than the wild-type protein for JH III (26). The Met gene was cloned and found to be a member of the basic helix-loop-helix-PER-AHR/ARNT-SIM (PAS) family of transcr...

DNA markers were identified for the molecular detection of the Asian long-horned beetle (ALB), Anoplophora glabripennis (Mot.), based on sequence characterized amplified regions (SCARs) derived from random amplified polymorphic DNA (RAPD) fragments. A 2,740-bp DNA fragment that was present only in ALB and not in other Cerambycids was identified after screening 230 random primers in a PCR-based assay system. Three pairs of nested 22-mer oligonucleotide primers were designed on the basis of the sequence of this fragment and were used to perform diagnostic PCR. The first pair of primers (SCAR1) amplified a single 745-bp fragment of ALB DNA, but this did not differentiate ALB from other species. The other two pairs of SCAR primers (SCAR2 and SCAR3) amplified bands of 1,237- and 2,720-bp, respectively, that were capable of differentiating ALB from other closely related non-native and native Cerambycids, such as A. chinensis (Forster), A. malasiaca (Thomson), A. nobilis (Ganglbauer), Monochamus scutellatus (Say), Plectrodera scalator (Fab), Saperda tridentata (Olivier), and Graphisurus fasciatus (Degeer). The latter two SCAR markers could be amplified using DNA extracted from body parts of ALB such as the wing, the leg, and the antennae as well as tissues from all the developmental stages including the egg, larva, pupa, and adult. These markers were also capable of identifying ALB using the DNA extracted from frass. Our results demonstrate that the SCAR markers we have identified can be used for unambiguously identifying ALB from other closely related Cerambycids using a simple PCR procedure.