Analysis of a mosquito acetylcholinesterase gene promoter

Abstract

Insect acetylcholinesterase is the target site for organophosphorus and carbamate insecticides and point mutations in the Ace gene are associated with resistance in Drosophila melanogaster and Musca domestica. However, little is known of the genetic regulation of insect Ace genes. Here we report the isolation of four different cDNAs from an Aedes Ace locus and identification of the gene promoter. Northern analysis reveals two large (>10 kb) transcripts and one smaller transcript of 4 kb. The region containing the initiation of transcription was localized by sequencing the two 5' most cDNAs and by 5' RACE. The transcription start point was subsequently identified by primer extension and is flanked by a perfect arthropod initiator consensus sequence. The promoter lacks a TATA box but contains several matches to other consensus sequences for eukaryotic transcription factors. In common with the Drosophila Ace gene, there are also multiple potential initiators of translation (ATGs) upstream of the main open reading frame. The structure of the 5' leader and promoter is compared to that found in other insect and vertebrate Ace genes and the possibility that this locus is homologous to one of two Ace loci described in another mosquito, Culex pipiens, is discussed.

Structure and promoter activity of the 5′ flanking region of ace-1, the gene encoding acetylcholinesterase of class A in Caenorhabditis elegans

“…In the absence of a TATA box, transcription may be initiated at multiple, tightly clustered sites that follow an initiator (Inr) consensus sequence (Smale & Baltimore, 1989). In insects, Inr sequences were also reported as functional promoter elements (Cherbas & Cherbas, 1993;Liu et al, 1998). Some of the transcription starts of ace-1 were found in sequences resembling an Inr consensus sequence (Javahery et al, 1994), but we did not test the functionality of these sequences in binding the preinitiation complex of transcription.…”

Genomic organization and immune regulation of the defensin gene from the mosquito, Anopheles gambiae

“…gambiae ribosomal protein gene (rpL7a) found no evidence of immune regulatory motifs of the kind described here (data not shown). Such elements are also absent from the promoters of the Aedes acetylcholinesterase and gamma‐aminobutyric acid receptor genes ( Liu et al. , 1998 ; Shotkoski et al.…”

Sensitive Raman spectroscopy of lipids based on drop deposition using DCDR and SERS

“…Moreover, cationic lipids and liposomes have also been widely used as transfection vehicles to introduce a gene inside the cells, and in vivo cationic liposome-mediated transfection requires low detection limits of liposomes. [26] Experimental Synthetic DMTAP (chloride salt, M.W.=590.361) in powder (Avanti Polar Lipids, Inc.) was dissolved in pure chloroform. A weak jet of nitrogen gas was used to eliminate chloroform and to form a thin lipid film at the bottom of the flask.…”