Open AccessRapid "footprinting" on supercoiled DNA.
Author(s) -
Jay D. Gralla
Publication year - 1985
Publication title -
proceedings of the national academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.82.10.3078
Subject(s) - dna supercoil , lac repressor , microbiology and biotechnology , polymerase , dna , dna footprinting , dna polymerase ii , dna polymerase , oligonucleotide , biology , dna clamp , transcription bubble , primase , chemistry , lac operon , biochemistry , rna , plasmid , dna replication , rna dependent rna polymerase , promoter , gene , reverse transcriptase , gene expression
A DNase protection technique is described and applied to the interaction of three lac control proteins with supercoiled lac DNA. The technique uses end-labeled oligonucleotide primers to probe specific DNA regions as an alternative to protocols requiring restriction endonuclease cleavage or blotting. Thus DNA may be probed with high resolution in its native state. It is demonstrated that the introduction of supercoiling into DNA accelerates the rate of lac ps promoter binding by RNA polymerase but does not alter the positions at which polymerase, c-AMP-binding protein, or lac repressor bind to lac DNA.
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