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Figure 1


Fig. 1. Disruption of the DYH7 gene. A 2.4 kb DNA fragment near the 5' end of the 14.5 kb DYH7 open reading frame (ORF) was cloned to pCR4-TOPO vector. The neo3 gene disruption cassette was inserted into ClaI and XmaI restriction sites that were introduced to this DYH7 fragment by site-directed mutagenesis. This disruption interrupts the predicted N-terminal tethering domain of DYH7 and is upstream of its P-loop domains. The resulting plasmid was linearized and used in biolistic transformation of vegetative CU427.4 Tetrahymena. The annealing sites for the DYH7 gene-specific primers (gsp) and neo3 cassette-specific primer (neo3BTU) are indicated.