Figure 3

Schematic representation of the « modular » PRIG vectors. pPRIGp ER is a pPRIG derivative containing the C-terminal portion of the mouse estrogen receptor bearing the G525V mutation. This mutant does not bind estradiol but still binds the synthetic ligand hydroxytamoxifene. The ER sequence has been cloned at the 3' end of the MCS and the reading frame is thus open from and throughout the MCS to the stop codon ending the ER sequence. In contrast, in pPRIGp VP16HA and pPRIGp KRABHA, the sequence coding the functional module (VP16 transactivating domain from human herpes virus or KRAB transrepressing domain from human KOX1 protein) was cloned upstream of the MCS. The reading frame is thus open from the start codon of the module to the stop codon of the in-frame HA sequence 3' of the MCS. Pa: site for PacI. The two PacI sites of pPRIGp VP16HA and pPRIGp KRABHA are symbolized by vertical bars. For each vector, the unique sites of the MCS are indicated above the MCS.