pcat (registered trademark) 3-enhancer  (Promega)

Journal: Gene Expression

Article Title: Homology between α2(V) and αl(lll) collagen promoters and evidence for negatively acting elements in the α2(V) first intron and 5′ flanking sequences

doi:

Figure Lengend Snippet: Promoter activities of sequences from the 5′ end of the human α2(V) gene. Arrows, beneath a partial restriction map of the 5′ portion of the α2(V) gene, denote the extent and orientation of α2(V) DNA fragments fused to the CAT gene for expression analysis. Bold arrows represent sequences, from −151 to +82, found to have strongest promoter activity. CAT activity of each plasmid is expressed relative to the conversion level achieved by the vector pCAT-control (100.0). pCAT-control (Promega, technical bulletin #81) is similar in configuration to pCAT Enhancer but contains a 200 bp Sphl-HindII fragment (128–5171) containing the SV40 early promoter, upstream of the CAT gene. (+) and (−) designate the presence or absence, respectively, of the SV40 enhancer downstream of the CAT gene in various plasmids. Data represent means ± standard deviation (in parenthesis) of from four to six independent transfection experiments. NA: not ascertained.

Article Snippet: CAT activity of each plasmid is expressed relative to the conversion level achieved by the vector pCAT-control (100.0). pCAT-control (Promega, technical bulletin #81) is similar in configuration to pCAT Enhancer but contains a 200 bp Sphl-HindII fragment (128–5171) containing the SV40 early promoter, upstream of the CAT gene. (+) and (−) designate the presence or absence, respectively, of the SV40 enhancer downstream of the CAT gene in various plasmids.

Techniques: Expressing, Activity Assay, Plasmid Preparation, Standard Deviation, Transfection

Journal: Immune Network

Article Title: A New Reporter Vector System Based on Flow-Cytometry to Detect Promoter Activity

doi: 10.4110/in.2009.9.6.243

Figure Lengend Snippet: Comparison of the dual reporter system with the conventional CAT reporter system. The same promoter regions used in the dual reporter vector system were re-cloned into the CAT reporter system, pCAT®3-Enhancer vector (A). Vectors harboring different lengths of the promoter region were cotransfected with pSV-β-Galactosidase vector. The transcriptional activities of each region were determined via Chloramphenicol acetyl transferase activity assays (B, C) after compensation with beta-galactosidase. The pCAT3 control vector without the promoter sequence (pCAT®3-Basic) and the one harboring the SV40 promoter (pCAT®3-Control) were used as negative and positive controls, respectively.

Article Snippet: Several promoter regions used in the dual reporter vector were re-cloned into the CAT expression vector system pCAT®3-Enhancer vector (Promega, Madison, WI), in order to ascertain whether these regions evidenced similar transcription activity in the CAT assay system ( ).

Techniques: Plasmid Preparation, Clone Assay, Chloramphenicol Acetyltransferase Assay, Activity Assay, Sequencing