Fig. 2. Characterisation of flp1::kanMX6 cells. (A) The flp1::kanMX6 allele structure. The location of the fragment used to probe the Southern blot shown in B is indicated, as are the positions of the EcoRI sites that give rise to the fragments observed in the autoradiograph. Correct replacement of the flp1⁺ gene gives rise to a 2.4 kb fragment. (B) Southern blot of DNA prepared from wild-type and flp1::kanMX6 haploid cells. Note the absence of the 3.3 kb band in the flp1::kanMX6 cells. (C,D) flp1::kanMX6 cells were grown in YE at 25°C to mid-exponential phase (C) and shifted to 36°C for 5 hours (D). (C) Cell 1 is a postmitotic, binucleate cell. (D) Cell 1 is multicompartmented; cell 2 has septated once, and then undergone at least one more mitosis without septating; cell 3 has septated centrally, but both nuclei are located on the same side of the septum; and cell 4 shows one of its nuclei bisected by the developing division septum.