Shotkoski F, Zhang H G, Jackson M B, ffrench-Constant R H
Department of Neuroscience, University of Wisconsin-Madison, 53706, USA.
FEBS Lett. 1996 Feb 19;380(3):257-62. doi: 10.1016/0014-5793(96)00025-7.
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.
我们致力于建立稳定转化的昆虫细胞系，高效表达昆虫γ-氨基丁酸（GABA）受体亚基基因——对狄氏剂有抗性或Rdl。为便于实现这一目标，我们利用了一个基于白纹伊蚊蚊子细胞系稳定转化的系统，该系统使用二氢叶酸还原酶（dhfr）基因作为选择标记。在此，我们报告了携带来自果蝇和埃及伊蚊蚊子的高拷贝数Rdl基因的稳定蚊子细胞系的产生，以及随后功能性GABA门控氯离子通道的高效表达。我们还利用该系统比较了一系列杆状病毒立即早期启动子在蚊子细胞培养中的活性，并证明IE1启动子构建体在跨昆虫物种中有效发挥作用。讨论了这些构建体在非果蝇类昆虫的通用转化中的潜在用途的相关结果。
