Authors
Tom Ellis
Summary
Parallel-synthesis of biomolecular components offers a rapid path to constructing multiple gene networks that behave as predicted by in silico modeling. Here we describe the synthesis and screening steps to produce a component library, using the example of repressible promoters for S. cerevisiae. Regulatory promoters such as these are crucial components in genetic networks and quantified variability in input and output can be used to design diverse networks. The parallel synthesis approach described here is not limited to promoters or S. cerevisiae and should be broadly applicable to all biomolecular components with protocol adaptations. Three main methods exist for producing component libraries; (i) combinatorial shuffling (Cox et al., 2007; Gertz et al., 2009; Ligr et al., 2006; Murphy et al., 2007), (ii) multi-round mutation/selection of an existing component (Alper et al., 2005; Alper et al., 2006), and (iii) diversity-inherent component synthesis (Jensen and Hammer, 1998a, b). We describe here the third method, which we recommend as it is rapid and compatible with rational design of new components.Further details
The protocol was published on Protocol Exchange on 23 April 2009. To see the entire protocol, click on the source link.Advertisement
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