Team:WITS-South Africa/Machine Design

Machine Design

Ideal Machines

Lacto-Detect

Lacto-Report

Intermediate Machines

etc

Machine Construction

Machine Testing

Final Machine 1 - LactoDetect

Purpose:

a) To act as the ‘Detector’ Machine within the population and produce the quorum signalling peptide in response to an input signal

Construction: LactoDetect was contructed via a series of cloning steps using the standard assembly method. These are outlined in Figure 1 below.

Intermediate Machine 2 (Figure 1) Purpose: a) To characterise the strength of the induction of the LacI/AraC promoter by measuring fluorescence after IPTG is added b) To show that the PlcR promoter is activated in L. gasseri by measuring fluorescence after the addition of exogenous PlcR and PapR proteins

Fig1. The proposed intermediate Machine 2

Fig 2. The proposed Final Machine 1

Final Machine 2 (Figure 3) Purpose a) To act as the ‘Reporter’ Machine within the population and respond to infection as previously described

Fig 3. The proposed Final Machine 2