Difference between revisions of "Part:BBa K831009"
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=='''Functional characterization'''== | =='''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). | + | 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). |
[[Image:BBa_K831009_1.png|center|550 px]] | [[Image:BBa_K831009_1.png|center|550 px]] | ||
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. | 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. | ||
Revision as of 00:34, 1 October 2012
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 223
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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).
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.