Fig. 2.
The deficiencies of 1C12 and 45G7 flagella were caused by mutations in the RSP6 gene. (A) A schematic diagram depicting the mutations in the RSP6 genes in three RSP6 mutants. For 1C12, a single-nucleotide deletion in the first exon resulted in premature termination after 58 amino acids. In the case of 45G7, insertion of pMN24 plasmid containing nitrate reductase gene (NIT1) in the fifth intron resulted in deletion of three fifth of the 3' end. An addition of one base pair in the last exon in the pf26ts RSP6 gene caused frame shift of the C-terminus and the presence of the mutated RSP6 in pf26 axoneme (Huang et al., 1981). The results were obtained by PCR of genomic DNA, followed by cloning in some cases and sequencing. The mutations were confirmed by repeated PCR and sequencing. The black bars represent exons and the two gray bars represent 5' and 3' UTR respectively. The lines between black bars represent introns. (B) Western analyses revealed that RSP6 was restored in two representative transformants (H4 and H5) to the WT level. The positive and negative controls were WT and 1C12 respectively. The protein loading control was dynein intermediate chain, IC140.