Part:BBa_K4090008

AraC-pBAD-Kid

Introduction

AraC-pBAD is the inducible system, in which Arabinose is the inducer. Kid toxin is a kind of toxin that E.coli produced by itself. When it is over expressed, it could kill the E.coli with high specificity. As a result, this biobrick is an useful kill switch of E.coli. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 160
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Viability

We designed the kill switch part as below

Fig.1

After transforming the plasmids into competent BL21 cells, we inoculated the cells in eight groups, including four in liquid medium and the other four in solid medium. The results are shown in the figures below. Arabinose was added in experimental groups. We tested and calculated the mean OD value of four liquid groups.

Fig.2

As shown in Fig.1-2, in the upper-left area of the medium, normal E.coli without the transformation of the plasmids could grow normally; in the upper-right area, transformed cells could grow normally without the presence of the Arabinose; in the bottom area, the cells were killed in the Arabinose-contained environment. Fig.1-3 shows that the OD value of liquid medium after adding Arabinose is lower than that of medium without Arabinose. These results indicate that this biobrick performed as effectively as expected.

Fig.3

Result of BN063 kill switch:

Reference

[1] Structural and Functional Analysis of the Kid Toxin Protein from E. coli Plasmid R1. Structure, Volume 10, Issue 10, October 2002, Pages 1425-1433.