FISH on Insect Cells Transfected with Heterologous DNA

Abstract

Insect cells transfected with heterologous DNAs (plasmids) can be used for the assessment of functional properties of genes that have been isolated or subjected to in vitro mutagenesis (Rubin and Spradling 1983, Snow et al. 1989). An example of a cell system used for such approaches are Schneider cells (Schneider 1972). These are derived from embryonic cells of Drosophila melanogaster, can be transfected with special plasmids (Bunch et al. 1988) and used for heterologous expression. Several, in general, reliable in vitro methods can be performed to control the transfection efficiency in such cells (i.e. PCR, Southern-, Northern- or Western-blot approaches). Another fast and reliable method to detect transfected Schneider cells, offering the possibility of assessing the transfection rate on a single cell level, has recently been described and is based on a modified FISH technique (Rautenstrauss et al. 1998; see Figure 1) and described in the following.