Dictyostelium discoideum has a number of features that make it an attractive system for cell biological studies. The ability of Dictyostelium cells to perform active ameboid crawling and chemotaxis has made it a popular system for cell motility and signal transduction studies. The lack of a cell wall and ease of cultivation make Dictyostelium an excellent organism for biochemical approaches, allowing large quantities of material to be obtained and lysed without difficulty. This chapter describes molecular genetic tools that are used for transformation, construction of null cell lines, and expression of the cloned myosin gene fragments. Brief coverage is given to other tools and methods common in the field, but the emphasis is on those approaches that are currently being used. Straightforward gene disruption protocols have been developed and used to construct myosin null lines of Dictyostelium. These techniques have also been successfully applied to the study of other genes in Dictyostelium, such as the α-actinin gene.
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