Combinatorial Synthesis of Genetic Networks
Author:
Affiliation:
1. Howard Hughes Medical Institute, Department of Molecular Biology,
2. The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.
3. Department of Physics, Princeton University, Princeton, NJ 08544, USA.
Abstract
Publisher
American Association for the Advancement of Science (AAAS)
Subject
Multidisciplinary
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4. In order to avoid toxicity effects due to overexpression and reductions in the dynamic regulatory range we used the reduced-stability variants of these transcriptional regulators described previously (21).
5. The following promoters were used. Poid70.5 (designated P L 2 ) from plasmid pOd O id 70.5 O 1 (22); P R (P λ - ) and P RM (P λ + ) from lambda phage DNA and P L lacO1 (P L 1 ) and P L tetO1 (P T ) from their original plasmids (23). Each of these promoters was amplified by polymerase chain reaction (PCR). In the case of P λ + (P RM ) the or3-r3 mutation (24) which eliminates repression of P RM at high λ CI concentrations was introduced by PCR.
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