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Barnase Growth Control

The fatal gene is the key element of the genetic fence, without which no containment will result. To confirm the ability of barnase to contain plants outside the genetic fence, we characterized an inducible barnase system created by the Berkeley 2008 iGEM team.

In this construct, arabinose induction results in Barnase expression. Because of the presence of low background levels of the inhibitor Barstar, low levels of Barnase expression will not immediately kill the cells. Higher and higher levels of Barnase expression results in lower and lower rates of growth in the cells, affirming the principle of Barnase-based growth control in the genetic fence.

Barnase Growth Control in E.Coli   click to enlarge

This barnase system works by expressing background levels of Barstar with Barnase inducible such that it will overwhelm Barstar when induced. A plate reader assay demonstrated the ability of Barnase to inhibit growth in Bacteria, confirming the results from Berkeley 2008.

Barstar Inhibition of Barnase