Characterization of promoter containing DNA fragments based on the abortive initiation reaction of Escherichia coli RNA polymerase.
The abortive initiation reaction of Escherichia coli RNA polymerase was demonstrated to be a general method for the rapid identification and quantitation of promoter sites on DNA. The presence of the T7 promoters, A1, A2, A3, and D on an isolated restriction fragment of the phage template was demonstrated. In addition, abortive initiation results indicated that the D promoter transcript started with pppGpUpUpG. This technique should prove particularly useful for screening DNA fragments for the number and type of promoters present.
D004279:DNA, Viral; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D008854:Microscopy, Electron; D009876:Operon; D011485:Protein Binding; D013604:T-Phages; D014158:Transcription, Genetic
D004279:DNA, Viral; D012321:DNA-Directed RNA Polymerases