Combinatorial Synthesis of Genetic Networks-Reference-Cited by-同舟云学术

Combinatorial Synthesis of Genetic Networks

Published:2002-05-24 Issue:5572 Volume:296 Page:1466-1470
ISSN:0036-8075
Container-title:Science
language:en
Short-container-title:Science

Author:

Guet Călin C.¹²,Elowitz Michael B.²,Hsing Weihong¹,Leibler Stanislas¹³²

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

A central problem in biology is determining how genes interact as parts of functional networks. Creation and analysis of synthetic networks, composed of well-characterized genetic elements, provide a framework for theoretical modeling. Here, with the use of a combinatorial method, a library of networks with varying connectivity was generated in Escherichia coli . These networks were composed of genes encoding the transcriptional regulators LacI, TetR, and lambda CI, as well as the corresponding promoters. They displayed phenotypic behaviors resembling binary logical circuits, with two chemical “inputs” and a fluorescent protein “output.” Within this simple system, diverse computational functions arose through changes in network connectivity. Combinatorial synthesis provides an alternative approach for studying biological networks, as well as an efficient method for producing diverse phenotypes in vivo.

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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