Figure 6

Injection of α1-GFPtel^- into the developing macronucleus of the polytene chromosome stage. The presence of the injected construct in the macronuclear DNA of injected cells and its processing was analyzed by PCR. After injection, the cells were allowed to finish macronuclear development, then a PCR analysis was performed using DNA of 30-40 vegetatively growing cells as templates. Primers used were P27, derived from the endogenous α1-tubulin macronuclear molecule, and pGFPP2, derived from GFP sequences. (a) Schematic diagram of the α1-GFPtel^- construct. (b) PCR products from the construct, from uninjected and injected cells were separated on an 1% agarose gel. M, molecular weight marker (1 kb ladder, Gibco, BRL). Lane 1, PCR product from construct DNA; lane 2, PCR product from uninjected cells; lane 3, PCR product from cells injected with α1-GFPtel^-. The gel was hybridized with a DIG-labeled probe of the α1-GFPtel^- vector DNA.