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Fig. 1. Isolation of MEL cells with large transgene arrays, tagged with lac operator repeats, and carrying ß-globin regulatory sequences. (A) The pPALZ8.8 plasmid was created by cloning several parts into pSV2-neo (black and dark green). Dotted blue and green boxes (flanked by ClaI and EcoRI restriction sites) indicate the hypersensitive sites HS1-HS4 of the human ß-globin locus which form the µLCR. The human ß-globin promoter (filled pink) controls a ß-galactosidase gene (LacZ, hollow pink). The light blue box indicates 64 repeats of the lac operator, the binding site for the GFP/lac-repressor fusion protein. Arrows indicate open reading frames of the genes involved. Total size of plasmid is 15 kb. Restriction sites mentioned in the Materials and Methods section are indicated. Sites in brackets had been present in the polylinker and were lost during cloning. (B) Cloning scheme for PALZ39 subclones.





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