The phage Mu middle promoter Pm contains a partial UP element.
Bottom Line: The results demonstrated that mutations upstream of -57 had no effect on Pm activity in vivo, assayed by expression of lacZ fused downstream of a wild-type or mutant Pm.In DNase I footprinting and gel mobility shift assays, paired mutations at positions -55 and -54 did not affect Mor binding but decreased the synergistic binding of Mor with histidine tagged α (His-α), indicating that His-α binds to Pm in a sequence- and/or structure-specific manner.Taken together, these results demonstrate that Pm has a strong proximal UP element subsite, but lacks a distal subsite.
Affiliation: Department of Microbiology, Immunology & Biochemistry, University of Tennessee Health Science Center, Memphis, Tennessee 38163.Show MeSH
Related in: MedlinePlus
Mentions: The promoter cloning plasmid, pIA12 (Figure 2), contains a HindIII-EcoRI-BamHI linker upstream from a promoter-less lacZ gene (Chiang and Howe 1993; Artsimovitch and Howe 1996). Plasmid pMM1 contains Pm sequence from −98 to +10 cloned into the EcoRI and BamHI sites in pIA12 (Chiang et al. 1993). Plasmid pKM78 (Figure 2) expresses Mor under the control of PlacUV5 (Mathee and Howe 1990). Plasmid pKM90 contains the mor gene downstream of the strong T7 promoter and ribosome binding site in pT7-7 (Tabor and Richardson 1985; Mathee and Howe 1993; Ma and Howe 2004). Plasmid pHTT7f1-NHα contains an rpoA gene encoding the N-terminal His6-tagged α subunit of E. coli RNA polymerase (Tang et al. 1994).
Affiliation: Department of Microbiology, Immunology & Biochemistry, University of Tennessee Health Science Center, Memphis, Tennessee 38163.