Part:BBa_K1467102:Experience

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Applications of BBa_K1467102

NRP-UEA iGEM 2014

iGEM14_NRP-UEA-Norwich have used this part to express reporter proteins when the construct is in the presence of the TALE AvrBS3. The aim of this was to test our system in which the presence of the pathogen (in this case the TALE AvrBS3) switches on the BS3 promoter, driving transcription of the coding sequence and therefore expression of the reporter protein of interest. These reporter proteins included GFP, Bax, NbHB1, AmilCP and AmilGFP. In constructs containing the promoter independent of AvrBS3, no transcription of the reporter protein occurred giving evidence that this part functions as expected. Examples of the promoter inducing GFP expression are shown below.

Figure 1: A Nicotiana benthamiana leaf under UV light following infiltration (red circles) with Agrobacterium tumefaciens carrying plasmids encoding BS3_GFP_ocs (B) and BS3_GFP_ocs + 35s_AvrBS3_nos (A) respectively.

There is GFP expression in the area infiltrated with the multigene construct containing both the BS3 promoter and the TALE AvrBS3, showing that the promoter is active (A). In the area in which the infiltrated agrobacterium was just carrying the transcriptional unit encoding the promoter, GFP and terminator, there is no expression of GFP as the BS3 promoter is not active (B). This shows the function of AvrBS3 in activating the inducible BS3 promoter.

User Reviews

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