An in vitro transposon system for highly regulated gene expression: construction of Escherichia coli strains with arabinose-dependent growth at low temperatures.

Placing a gene of interest under the control of an inducible promoter greatly aids the purification, localization and functional analysis of proteins but usually requires the sub-cloning of the gene of interest into an appropriate expression vector. Here, we describe an alternative approach employing in vitro transposition of Tn Omega P(BAD) to place the highly regulable, arabinose inducible P(BAD) promoter upstream of the gene to be expressed. The method is rapid, simple and facilitates the optimization of expression by producing constructs with variable distances between the P(BAD) promoter and the gene. To illustrate the use of this approach, we describe the construction of a strain of Escherichia coli in which growth at low temperatures on solid media is dependent on threshold levels of arabinose. Other uses of the transposable promoter are also discussed.

Knowledge modelling in aerospace design

This paper addresses the need for computer support in aerospace design. A review of current design methodologies and computer support tools is presented and the need for further support in aerospace design, particularly in the early formative stages of the design process, is discussed. A parameter-based model of design, founded on the assumption that a design process can be constructed from a predefined set of tasks, is proposed for aerospace design. This is supported by knowledge of possible tasks in which the confidence in key design parameters is used as a basis for identifying, or signposting, the next task. A prototype implementation of the signposting model, for use in the design of helicopter rotor blades, is described and results from trials of the tool are presented. Further areas of research are discussed