Difference between revisions of "Part:BBa K518010"
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<partinfo>BBa_K518010 short</partinfo> | <partinfo>BBa_K518010 short</partinfo> | ||
| − | Microbes including Escherichia coli are known to respond to various DNA-injuring stress (ionizing radiation, ultraviolet radiation, peroxides etc...), altering their gene | + | Microbes including ''Escherichia coli'' are known to respond to various DNA-injuring stress (ionizing radiation, ultraviolet radiation, peroxides etc...), altering their gene expression patterns. This response, known as the "SOS response", is induced by a regulatory protein called RecA when it binds to single-strand DNA. The DNA-RecA complex promotes the degradation of LexA, a common repressor of SOS genes. |
| − | SulA is responsible for stress-induced halt of cell division. The promoter of ''sulA'', | + | SulA is responsible for stress-induced halt of cell division. The promoter of ''sulA'', sulAp, is induced by various stress factors, including ultraviolet irradiation. |
===Application=== | ===Application=== | ||
| − | We utilized | + | We utilized this property (induced-expression by UV irradiation) to design a "UV switch". This makes it possible to "switch on" a genetic circuit using UV. As a first step for this, we characterized the UV-induction of sulAp. |
The UV-induced expression level of [[Part:BBa_K518010 |BBa_K518010]] was evaluated using [[Part:BBa_K518013 |BBa_K518013]]. As a measurement tool, our dual luciferase assay kit was employed. For detailed infomation, see [[Part:BBa_K518002 |BBa_K518002]]. | The UV-induced expression level of [[Part:BBa_K518010 |BBa_K518010]] was evaluated using [[Part:BBa_K518013 |BBa_K518013]]. As a measurement tool, our dual luciferase assay kit was employed. For detailed infomation, see [[Part:BBa_K518002 |BBa_K518002]]. | ||
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[[Image:sulApexpression.png]] | [[Image:sulApexpression.png]] | ||
| − | <Figure: UV-induced expression levels of [[Part:BBa_K518010 |sulAp]]. The expression levels | + | <Figure: UV-induced expression levels of[[Part:BBa_K518010 |sulAp]]. The expression levels from sulAp were evaluated before and after UV induction. We evaluated it using both ''recA''(-) (JM109) and ''recA''(+) (BL21) strain. RecA is known to be necessary for releasing sulAp from repression. We successfully demonstrated a significant alteration of expression in ''recA''(+) strain only after UV irradiation. The expression level of [[Part:BBa_J23119 |BBa_J23119]], a constitutive E. coli promoter which is often used as a comparison, was simultaneously presented. Data is expressed as mean ± S.D.. Data is obtained from the average of three independent experiments.> |
Revision as of 20:02, 5 October 2011
sulA promoter
Microbes including Escherichia coli are known to respond to various DNA-injuring stress (ionizing radiation, ultraviolet radiation, peroxides etc...), altering their gene expression patterns. This response, known as the "SOS response", is induced by a regulatory protein called RecA when it binds to single-strand DNA. The DNA-RecA complex promotes the degradation of LexA, a common repressor of SOS genes.
SulA is responsible for stress-induced halt of cell division. The promoter of sulA, sulAp, is induced by various stress factors, including ultraviolet irradiation.
Application
We utilized this property (induced-expression by UV irradiation) to design a "UV switch". This makes it possible to "switch on" a genetic circuit using UV. As a first step for this, we characterized the UV-induction of sulAp.
The UV-induced expression level of BBa_K518010 was evaluated using BBa_K518013. As a measurement tool, our dual luciferase assay kit was employed. For detailed infomation, see BBa_K518002.
<Figure: UV-induced expression levels ofsulAp. The expression levels from sulAp were evaluated before and after UV induction. We evaluated it using both recA(-) (JM109) and recA(+) (BL21) strain. RecA is known to be necessary for releasing sulAp from repression. We successfully demonstrated a significant alteration of expression in recA(+) strain only after UV irradiation. The expression level of BBa_J23119, a constitutive E. coli promoter which is often used as a comparison, was simultaneously presented. Data is expressed as mean ± S.D.. Data is obtained from the average of three independent experiments.>
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]