EXISTING BIOBRICK CHARACTERISATION

As part of the criterion for the Gold medal, we set about trying to improve the characterisation of an existing part on the registy. In line with the rest of our project, we felt it pertinent that we characterise something relevant and as such we chose this part. The information below is taken from the registry page:

Introduction

Warwick iGEM 2014 used this part as a positive control for our experiments investigating promoter strengths for a self replicating RNA strand, using the Hepatitis C Virus NS5B gene encoding RNA dependent RNA polymerase, as the replicating enzyme.

This was measured using a tecan Magellan plate reader in which fluorescence in the GFP range was measured over a period of 24 hours with both biological and technical replicates for each transformed cell type.

Experimental Design

Test: GFP expression rate in cells co-transformed with plasmid that contains the promoter and a reversed GFP sequence and a plasmid containing the RdRP (Part:BBa_K1442100).

Positive Control: GFP expression rate in cells transformed with Biobrick plasmid that contains a gene for GFP with an inducible T7 promoter (Part:BBa_J04430). Promoter induction strength in response to different concentrations of added inducer (IPTG) was characterised.

Negative Control: GFP expression rate in cells co-transformed with the Promoter Testing Module and a plasmid containing a mutated version of the RdRP (Part:BBa_K1442101) that has been proven to be unable to direct successful replication of RNA.