Structure of a promotor on plasmid pMB9 derived from plasmid pSC101.
The DNA sequence of a 354 basepair EcoRI-HindIII fragment of plasmid pMB9 which has originally been derived from plasmid pSC101 has been resolved. This fragment contains a promoter for transcription directed towards the EcoRI site. Escherichia coli RNA polymerase binds to a region within the EcoRI-HindIII fragment which contains the heptamer 5' TATGGTG (132-126) and the duodecamer 5' TGATGAACATCA (158-147). Based on commonalities with other promotors these DNA sequences probably function as, respectively, "binding site" and "recognition site". Furthermore, this fragment harbours a translation reading frame free of nonsense codons and at about 25 basepairs from the indicated heptamer a nucleotide sequence which meets with the requirements for initiation of translation. By heteroduplex mapping it was shown that the EcoRI-HindIII fragment has been derived from a region near or within the origin of replication of pSC101. The copynumber of plasmids containing the EcoRI-HindIII fragment is two-fold lower than that of plasmids lacking this fragment. This effect might be related to the original function of this fragment on plasmid pSC101.
D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D009838:Oligodeoxyribonucleotides / Q000032:analysis; D009876:Operon; D010957:Plasmids; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic; D001618:beta-Lactamases / Q000378:metabolism
D004274:DNA, Recombinant; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D012321:DNA-Directed RNA Polymerases; D004262:DNA Restriction Enzymes; D001618:beta-Lactamases