Molecular biology in Dictyostelium: tools and applications.
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This chapter discusses that the Dictyostelium transformation system provides a tool to study gene regulation in Dictyostelium at the molecular level. The major factors influencing molecular biological techniques in Dictyostelium are its genome organization and DNA composition. Gene fusions carrying the promoter regions of developmentally regulated Dictyostelium genes are expressed and regulated in a manner similar to their genomic counterparts. Up to now, this has been shown for an actin gene, two discoidin genes, the Dictyostelium ras gene, and pst-cath, a prestalk-specific gene. The first major application of the transformation system is the mapping of promoters and sequences essential for regulated gene expression. Trans-acting factors are investigated using cotransformation. It is possible that extrachromosomal vectors containing the promoter of a differentially regulated gene can be isolated at time points during development while maintaining their chromatin structure. The investigation of the protein organization in the promoter region on such a DNA gives insight into the DNA protein interactions during gene regulation. The antisense transformation provides means to study the phenotype of cells deficient in a known gene product, thus mimicking a loss of function mutant. 1987, Elsevier Inc. All rights reserved.
