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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
Authors:Frank Shotkoski  Hai-Guang Zhang  Meyer B Jackson  Richard H ffrench-Constant
Institution:aDepartments of Neuroscience and Entomology, University of Wisconsin-Madison, 237 Russell Laboratories, 1630 Linden Drive, Madison, WI 53706, USA;bDepartment of Physiology, University of Wisconsin-Madison, Madison, WI 53706, USA
Abstract:We are interested in establishing stably transformed insect cell lines efficiently expressing the insect γ-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 selectable 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 chloride 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 genetic transformation of non-Drosophilid insects.
Keywords:GABA receptor  Stable transformation  Cell culture  Aedes albopictus  Gene expression  Immediate early baculovirus promoter
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