From 2010.igem.org
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| | Wild type ''Bacillus subtillis'' | | | Wild type ''Bacillus subtillis'' |
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- | |[[Image:Wild type Bacillus subtillis.jpg]] | + | |[[Image:Wild type Bacillus subtillis.jpg|right]] |
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| |''dinR'' KO | | |''dinR'' KO |
Revision as of 08:42, 13 August 2010
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Molecular tweezer tensile strength test.
YneA
SOS response is believe to be a universal bacteria phenomenon first studied in E.coli -LexA, recA
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In Bacillus subtillis (gram positive) dinR protein is homologous to lexA (Repressor of din-damage inducible genes). din genes include uvrA, uvrB, dinB, dinC dinR and recA. DNA damage inhibits cell division.
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Wild type Bacillus subtillis
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dinR KO
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dinR KO mutant over expressed the divergent (opposite direction) transcript for YneA, YneB and YnzC. These genes form the SOS regulon (recA independent SOS response)
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YneA suppressed in wt without SOS induction #
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Expression of YneA from IPTG controlled promoter in wt leads to elongation.
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Disruption of YneA in SOS response leads to reduced elongation. Altering YneB and YnzC expression does not affect cell morphology.
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Double mutant (dinR YneA)
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YneA protein required to suppress cell division. Not chromosome replication or segregation.
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FtsZ is important for bacterial cell division forming a ring structure at the division site by polymerising
assembling other proteins necessary for division at the site.
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FtsZ localises to the cell division cycle unless dinR is disrupted or YneA is being induced.
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YneA suppresses FtsZ ring formation- no proven direct interaction by two-hybrid.
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Filamentous cells less colony formation.
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YneA expression via the inactivation of dinR by Rec A is important.
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Sequence of YneA: http://www.ncbi.nlm.nih.gov/nuccore/NC_000964.3?from=1918391&to=1918738&report=graph&content=5
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Kawai, Y., Moriya, S., & Ogasawara, N. (2003). Identification of a protein, YneA, responsible for cell division suppression during the SOS response in Bacillus subtilis. Molecular microbiology, 47(4), 1113-22. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/12581363.
yneA cloning strategy
Media:yneA cloning strategy.pdf