A matched set of cat vectors for rapid mutational analysis of eukaryotic promoters and enhancers.
The eukaryotic cat expression vectors, pBRAMScat1 and pBRAMScat2, were constructed to simplify the analysis of genomic fragments containing putative transcriptional regulatory elements. These vectors contain the f1 filamentous phage origin of replication for single-stranded DNA rescue, and permit site-directed mutagenesis, and dideoxy sequencing of nested deletion mutants using commercial T3, T7 and M13 universal forward/reverse primers. The above features eliminate the need to shuttle back and forth between a conventional cloning vector and the cat expression vector during the analysis of putative eukaryotic gene regulatory elements. Plasmid pBRAMScat1 contains the bacterial chloramphenicol acetyltransferase-encoding gene (cat) and no eukaryotic promoter and was designed for the analysis of eukaryotic promoters. Plasmid pBRAMScat2 contains the cat gene under the control of the Herpes simplex virus thymidine kinase promoter and was designed for the analysis of eukaryotic enhancers.
Iyer
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Molecular Immunology Laboratory, Massachusetts General Hospital, Charlestown.
D000818:Animals; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D018139:Simplexvirus / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics
D009838:Oligodeoxyribonucleotides; D015500:Chloramphenicol O-Acetyltransferase; D013937:Thymidine Kinase