Part:BBa_K518013

sulA promoter evaluation device

Microbes including Escherichia coli are known to respond to various DNA-injuring stress (ionizing radiation, ultraviolet radiation, peroxides etc...), altering their gene expressions. This response, known as "SOS response", are induced by regulatory protein called RecA, which is activated when binds to single-strand DNA. DNA-RecA complex promotes the self-degradation of LexA, a common repressor for SOS genes.

SulA is responsible for stress-induced halt of cell division. The promoter of sulA, or sulAp, is acutely induced by various stress factors, including ultraviolet irradiation.

We provide a sulAp evaluation device to make it easy to measure the varying expression of sulAp in various "stressful" conditions. Employing our dual luciferase assay kit, quantitative measurement and comparison of Relative Promoter Unit (RPU) is achieved.

Measurement

<Figure: UV-induced expression levels of sulAp. The expression levels of sulAp were evaluated before and after UV induction. We evaluated it using both recA(-) (JM109) and recA(+) (BL21) strain. RecA is known as necessary for releasing sulAp from repression. We successfully demonstrate a significant alteration of expression in recA(+) strain after UV irradiation. The expression level of BBa_J23119, a consensus sequence of E. coli promoters, was simultaneously presented as a comparison. Data is expressed as mean ± S.D.. Data is obtained from three independent experiments.>

For experimental details, see [http://2011.igem.org/Team:UT-Tokyo our page]. For detailed information on the dual luciferase assay kit, see K518002.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1850
  Illegal NheI site found at 1873
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2452
  Illegal SapI.rc site found at 887