Directed evolution: new parts and optimized function

Dougherty, Michael J.; Arnold, Frances H.

doi:10.1016/j.copbio.2009.08.005

Cited by 134 publications

(106 citation statements)

References 31 publications

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“…Directed evolution in the laboratory (in vitro) mimics natural evolution but operates at the molecular level (i.e. no new organism is created) and focuses on specific molecular properties (5).…”

Section: Introductionmentioning

confidence: 99%

Optimizing the binding activity of the AP2/ERF transcription factor with the GCC box element from Brassica napus by directed evolution

Jin

Zhu²,

Peng³

et al. 2010

BMB Reports

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In this study, we cloned the ERF-B3 subfamily transcription factor gene BnaERF-B3-hy15 from Brassica napus L. Huyou15. This 600 bp gene encodes a 199 amino acid classic ethylene responsive factor (ERF), which shown no binding or very weak binding GCC box-binding activity by the yeast one-hybrid assay. We used gene shuffling and the yeast one-hybrid system to obtain three mutated sequences that can bind to the GCC box. Sequence analysis indicated that two residues, Gly156 in the AP2 domain and Phe62 at the N-terminal domain were mutated to arginine and serine, respectively. Changes of Gly156 to arginine and Phe62 to serine increased the GCCbinding activity of BnaERF-B3-hy15 and the alter of Gly156 to arginine changed the AP2-domain structure of BnaERF-B3-hy15. [BMB reports 2010; 43(8): 567-572]

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Section: Introductionmentioning

confidence: 99%

Optimizing the binding activity of the AP2/ERF transcription factor with the GCC box element from Brassica napus by directed evolution

Jin

Zhu²,

Peng³

et al. 2010

BMB Reports

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“…The idea that other living organisms can be utilised for human purposes is probably as 5 Some synthetic biologists increasingly apply directed evolution as a non-rational designing aid (Dougherty and Arnold 2009). This constitutes an interesting withdrawal from the machine-analogy that might indicate that maybe in the end the rational design of a living organism might be beyond human capacities.…”

Section: Utilising Lifementioning

confidence: 99%

The Conception of Life in Synthetic Biology

Deplazes‐Zemp

2011

Sci Eng Ethics

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The phrase 'synthetic biology' is used to describe a set of different scientific and technological disciplines, which share the objective to design and produce new life forms. This essay addresses the following questions: What conception of life stands behind this ambitious objective? In what relation does this conception of life stand to that of traditional biology and biotechnology? And, could such a conception of life raise ethical concerns? Three different observations that provide useful indications for the conception of life in synthetic biology will be discussed in detail: 1. Synthetic biologists focus on different features of living organisms in order to design new life forms, 2. Synthetic biologists want to contribute to the understanding of life, and 3. Synthetic biologists want to modify life through a rational design, which implies the notions of utilising, minimising/optimising, varying and overcoming life. These observations indicate a tight connection between science and technology, a focus on selected aspects of life, a production-oriented approach to life, and a design-oriented understanding of life. It will be argued that through this conception of life synthetic biologists present life in a different light. This conception of life will be illustrated by the metaphor of a toolbox. According to the notion of life as a toolbox, the different features of living organisms are perceived as various rationally designed instruments that can be used for the production of the living organism itself or secondary products made by the organism. According to certain ethical positions this conception of life might raise ethical concerns related to the status of the organism, the motives of the scientists and the role of technology in our society. Synthetic biologists want to modify life through a rational design, which implies the notions of utilising, minimising/optimising, varying and overcoming life.These observations indicate a tight connection between science and technology, a focus on selected aspects of life, a production-oriented approach to life, and a designoriented understanding of life. It will be argued that through this conception of life synthetic biologists present life in a different light. This conception of life will be illustrated by the metaphor of a toolbox. According to the notion of life as a toolbox, the different features of living organisms are perceived as various rationally designed instruments that can be used for the production of the living organism itself or secondary products made by the organism. According to certain ethical positions this conception of life might raise ethical concerns related to the status of the organism, the motives of the scientists and the role of technology in our society.

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“…Directed evolution is an alternative method that can effectively address this limitation. Many synthetic biology applications will require parts for genetic circuits, cell-cell communication systems, and non-natural metabolic pathways that cannot be found in Nature, simply because Nature is not in need of them (Dougherty & Arnold, 2009). In essence, directed evolution begins with the generation of a library containing many different DNA molecules, often by error-prone DNA replication, DNA shuffling or combinatorial synthesis (Crameri et al, 1998).…”

Section: A Roadmap From Design To Production Of New Drugsmentioning

confidence: 99%