Pgl 4 basic luciferase expression vector

The Src promoter (−39713 to −39762, pSrcwt) was amplified from human blood genomic DNA (Clontech, Mountain View, California) using a forward primer containing a KpnI restriction enzyme cleavage site (5′-CGGGGTACCAAGAGGCTTGATTGGTAGCGTA-3′) and a reverse primer containing an XhoI restriction enzyme cleavage site (5′-CCGCTCGAGGCTCCAGGGCTATCCTTTTGGG-3′) and was cloned into the pGL4 basic luciferase expression vector (Promega). To generate an Src mutant that is deleted at the core binding site of PXN, -GATG-was deleted from the promoter region of Src. The QuickChange IIXL Site-Directed Mutagenesis Kit (Stratagene, La Jolla, California) was used to generate mutant Src (pSrcmut) using the SRC luciferase construct as a template and the following primers: forward primer 5′-CATTCTTGAATATGGAGTTGGGAGTTGGTTCTTGCAAGTAGG-3′ and reverse primer 5′-CCTACTTGCAAGAACCAACTCCCAACTCCATATTCAAGAATG-3′. These vectors were produced by the core for molecular cloning in the Department of Cancer Biology at MD Anderson. The PLAT promoter was amplified from human blood genomic DNA using a forward primer containing an XhoI restriction enzyme site (5′-GGCTCGAGTCTATTTCCAGGGGCATT-3′) and a reverse primer containing a Hind III restriction enzyme site (5′-GGAAGCTTGAGACAGACCCCAAGGTA-3′) and was cloned into the pGL4 basic luciferase expression vector.