Joerg Hoeflich et al. (2002) European Worm Meeting "Cloning and characterization of srf-3"

Sam Politz, Joerg Hoeflich, Jonathan Hodgkin, Ralf Baumeister

[European Worm Meeting, 2002]

The nematode cuticle is a complex collagenous structure which is important for maintenance of morphology and locomotion. The cuticle consists of at least two distinct classes of proteins: the collagens and the surface associated proteins. Mutations in collagens very often result in animals with altered morphology e.g. dpy or rol phenotype. Mutations affecting cuticle structure have been extensively studied in Caenorhabditis elegans and various dpy, rol and sqt mutations have been described in molecular detail.

Joerg Hoeflich et al. (2001) International C. elegans Meeting "Tetracycline-inducible gene expression system in Caenorhabditis elegans"

Joerg Hoeflich, Ralf Baumeister

[International C. elegans Meeting, 2001]

A significant portion of mutations in Caenorhabditis elegans genes cause lethality. To study late effects of a lethal mutant, an inducible promotor system is desireable. In addition, ectopic expression of a candidate gene is a useful way to learn about the (mis-)function of a gene. Ectopic expression in C. elegans can be achieved by using a heat shock inducible promotor or by using different endogenous promotors. However for most of these promotors the time of expression and the expression level cannot be controlled. In contrast, inducible promotor systems based on chimeric transcription factors have been widely used to regulate gene expression in many different eukaryotic systems (e.g. insects, mice, yeast and plants). In order to test the efficiency of the tetracycline (Tc) responsive regulatory system in C. elegans we generated lines expressing the tetracycline-controlled transactivator proteins (tTA2/rtTA2). These proteins bind to DNA and therefore activate transcription in the presence (rtTA2) or absence (tTA2) of the antibiotics tetracycline or doxycycline (a more potent tetracycline agonist). We have crossed these lines with lines containing a tTA2/rtTA2 responsive promotor fused to a reporter gene and showed that gene expression could be induced (rtTA2) or inhibited (tTA2) by doxycycline treatment. We are constructing different promotor-transactivator fusions to test the ability of this system to direct gene expression in specific cells.