Abstract
This chapter presents methods for the biolistic transformation of both macro- and micronuclei of Tetrahymena thermophila. DNA-coated gold particles are fired into starved vegetative cells and, during late phases of conjugation, into mating cells to introduce gene sequences into the macronucleus. Early stages of mating are bombarded to add genes to the micronucleus. Methods for cell preparation and a detailed protocol for shooting and recovering transformed cells are also presented. Development of successful transformation techniques in any ciliate is complicated by the separation of somatic and germinal functions into two distinctly different nuclei: the polyploid macronucleus, which solely directs the cell's phenotype, and the transcriptionally inert diploid micronucleus, which contributes the genotype to subsequent sexual generations. Each of these nuclei is a unique target for DNA transformation, requires different strategies for the introduction of genes, and responds in different ways following successful transformation.
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