title: Stable expression of insect GABA receptors in insect cell lines. Promoters for efficient expression of Drosophila and mosquito Rdl GABA receptors in stably transformed mosquito cell lines.
abstract: We are interested in establishing stably transformed insect cell lines efficiently expressing the insect gamma-aminobutyric acid (GABA) receptor subunit gene Resistance to dieldrin or Rdl. In order to facilitate this we utilized a system based on stable transformation of Aedes albopictus mosquito cell lines using the dihydrofolate reductase (dhfr) gene as a selected marker. Here we report the production of stable mosquito cell lines carrying high copy numbers of Rdl genes from both Drosophila and Aedes aegypti mosquitoes and the subsequent high efficiency expression of functional GABA gated chlorine ion channels. We also used this system to compare the activity of a range of immediate early baculovirus promoters in mosquito cell culture and demonstrate that IE1 promoter constructs work efficiently across insect species. Results are discussed in relation to the potential use of these constructs in the generic transformation of non-Drosophilid insects.
authors.affiliation: Department of Neuroscience, University of Wisconsin-Madison, 53706, USA.
authors.forename: F
authors.initials: F
authors.lastname: Shotkoski
authors.forename: H G
authors.initials: HG
authors.lastname: Zhang
authors.forename: M B
authors.initials: MB
authors.lastname: Jackson
authors.forename: R H
authors.initials: RH
authors.lastname: ffrench-Constant
mesh_terms: D000330:Aedes / Q000235:genetics*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002461:Cell Line, Transformed; D018118:Chloride Channels / Q000235:genetics; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins; D015870:Gene Expression; D005819:Genetic Markers; D007305:Insecticide Resistance / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011963:Receptors, GABA-A / Q000235:genetics*; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014162:Transfection
chemical_list: D018118:Chloride Channels; D029721:Drosophila Proteins; D005819:Genetic Markers; C082030:Rdl protein, Drosophila; D011963:Receptors, GABA-A; D013762:Tetrahydrofolate Dehydrogenase