Cloning and Characterizing CXCR4 and CXCR7 Promoters

The promoter region of the CXCR4 and CXCR7 genes, from -2,000 bp to +50 bp, relative to the transcription start site were amplified according to previous report [24 (link)]. The confirmed sequence was then inserted into a pGL3-Basic vector (Promega). The pGL3-Basic vector was digested by KpnI and XhoI (or XhoI /HindIII for CXCR7; all restriction enzymes were acquired from MBI Fermentas), and the amplified CXCR4 and CXCR7 promoter fragments were inserted through ligation. The cloned pGL3-CXCR4 and pGL3-CXCR7 constructs were confirmed by sequencing. The sequentially shorter CXCR4 and CXCR7 promoter fragments were amplified by standard PCR methods, sequenced, and cloned to the pGL3 vector.