Compositions and methods for determining the activity of DNA-binding proteins and of initiation of transcription
First Claim
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1. A recombinant plasmid comprising:
- (a) a first region comprising a nucleotide sequence which specifically binds to a sequence-specific DNA-binding molecule;
(b) a second region comprising a nucleotide sequence which binds to a restriction enzyme; and
(c) a restriction site for said restriction enzyme;
wherein said restriction site is located at a position selected from being within said first nucleotide sequence, within said second nucleotide sequence, substantially adjacent to said first nucleotide sequence, and substantially adjacent to said second nucleotide sequence.
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Abstract
This invention provides plasmids that are useful in detecting and determining the DNA-binding activity of sequence-specific DNA-binding molecules. The invention further provides plasmids that are useful in detecting and determining the activity of RNA polymerases in initiating transcription. In particular, the invention relates to plasmids that contain unique restriction sites and cognate nucleotide recognition sequences for sequence-specific DNA-binding molecules. Also provided are methods for using the plasmids disclosed herein.
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8 Claims
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1. A recombinant plasmid comprising:
- (a) a first region comprising a nucleotide sequence which specifically binds to a sequence-specific DNA-binding molecule;
(b) a second region comprising a nucleotide sequence which binds to a restriction enzyme; and
(c) a restriction site for said restriction enzyme;
wherein said restriction site is located at a position selected from being within said first nucleotide sequence, within said second nucleotide sequence, substantially adjacent to said first nucleotide sequence, and substantially adjacent to said second nucleotide sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
- (a) a first region comprising a nucleotide sequence which specifically binds to a sequence-specific DNA-binding molecule;
Specification