The gene mutated in ssh1 was determined by AFLP analysis (Kathir et al., 2003 (link)) in combination with the Chlamydomonas genome database (genome.jgi-psf.org/Chlre4/Chlre4.home.html) and flagella proteome database (Pazour et al., 2005 (link)). The primers used in this study are listed in Supplemental Table S2 of Kubo et al. (2014) (link).
To generate pf28pf30tpg1::TTLL9HA strains, pf28pf30tpg1 was cotransformed with a construct encoding TTLL9-HA (Kubo et al., 2014 (link)) and the paromomycin resistance gene and selected on a TAP agar plate containing 10 μg/ml paromomycin (Sigma, St. Louis, MO). Expression of TTLL9-HA was confirmed by Western blotting using anti–HA-tag antibody (3F10).
To generate pf28pf30tpg1::TTLL9HA strains, pf28pf30tpg1 was cotransformed with a construct encoding TTLL9-HA (Kubo et al., 2014 (link)) and the paromomycin resistance gene and selected on a TAP agar plate containing 10 μg/ml paromomycin (Sigma, St. Louis, MO). Expression of TTLL9-HA was confirmed by Western blotting using anti–HA-tag antibody (3F10).
