Figure 5

Flp-mediated deletion and resulting luciferase expression in Rht14-10IN5 and Rht14-19IN5. A. Ethidium bromide stained agarose gel of electrophoresed PCR products generated with the primers O2 and O4 (Fig. 4A) and cell pellets of Rht14-10, Rht14-10IN5, pTIGHTluc (+) and Rht10IN5flp pools (top, 1–16) or clones constituting Rht10IN5flp pools 5 and 9 (bottom). Positive controls (+) used pTIGHTluc DNA (expected product: 517 bp). B. Luciferase activity in lysates from clones indicated (*) in A, or control cells (Rh14-10), grown in the absence or the presence (for 48 h) of tet. C. Ethidium bromide stained agarose gel of electrophoresed PCR products generated with the primers O2 and O4 (Fig. 4A) and cell pellets of Rht14-19, Rht14-19IN5, and Rht19IN5flp pools (top, 1–12) or constituent clones of pools 9 and 12 (bottom, 1–6). Positive controls (+) were as in A. D. Luciferase activity in lysates from clones indicated (*) in C, or control cells (Rht14-19), grown in the absence or the presence (for 48 h) of tet. Results of two experiments are shown.