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Part:BBa_K831009

Designed by: Silvia J Caas   Group: iGEM12_Colombia   (2012-09-26)
Revision as of 00:34, 1 October 2012 by Sylvita1015 (Talk | contribs) (→‎Functional characterization)

Inducible HipB antitoxin under the control of lac promoter

HipB is the antitoxin of the HipAB toxin-antitoxin (TA) module. HipB neutralizes the toxin effect of HipA due to protein-protein interactions.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 223
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Functional characterization

As HipB neutralizes the induction of persistency caused by HipA7 in this strain, we induced our strain with IPTG at different concentrations and evaluate the persisters frequencies implementing an original protocol for persister cells isolation based on lysis (S. Cañas et al., Manuscript in preparation).

BBa K831009 1.png

For this experiment with used the Escherichia coli K12 TH1269 strain (hipA7) which is a high persistence mutant. We evaluated the persistence frequencies of untransformed TH1269 strain and Escherichia coli MG1655 controls. This experiment was made in LB medium, supplemented with Ampicillin (100 ug/mL) for ensuring plasmid maintenance, when cultures reached stationary growth phase (ON). Bacterial cultures were incubated at 37 C and 200 rpm.

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n/aInducible HipB antitoxin under the control of lac promoter