DNA rearrangements generating artificial promoters.
The promoter-cloning plasmid pBRH4 (a derivative of pBR322 with a partially deleted promoter of the tet gene) is shown to contain a sequence which is located near the EcoRI site and can operate as an effective Pribnow box, but is not the remainder of the deletion-inactivated tet promoter of pBR322. If there is a sequence homologous to the '-35' promoter region at the border of the DNA fragment inserted at the EcoRI site, then a compound promoter arises and activates the tet gene. Point mutations in the nonfunctional--35 region of pBRH4 also activate the cryptic Pribnow box. Several compound promoters were obtained through deleting small portions of DNA around the HindIII site of pBR322; the deletions moved various sequences that could operate as Pribnow boxes towards the -35 region of the tet promoter.
D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis*; D015246:Deoxyribonuclease EcoRI; D004720:Endonucleases / Q000378:metabolism; D009876:Operon; D010957:Plasmids; D015719:Single-Strand Specific DNA and RNA Endonucleases
D004269:DNA, Bacterial; D004720:Endonucleases; D004262:DNA Restriction Enzymes; D015246:Deoxyribonuclease EcoRI; D015719:Single-Strand Specific DNA and RNA Endonucleases