Difference between revisions of "Part:BBa K174008"

Revision as of 22:17, 21 October 2009

SmtA metallothionein protein with CotC and Gfp fusion

SmtA metallothionein protein from E. coli can bind to heavy metals [1,2,3]. Metallothioneins have a tendency to bind to cationic metal ions such as cadmium, copper, arsenic, mercury, silver.

By translationally fusing CotC spore coat protein from Bacillus subtilis, SmtA can be localized to the spore coat, hence trapping heavy metals into bacterial spores. SmtA is also fused with Gfp to facilitate the visualisation of spores using a fluorescent microscope.

We designed this device to sequester cadmium into B. subtilis spores. The device is expressed using a sigK type promoter which becomes active under sporulation conditions. Hence metals are soaked up when the cells are undergoing sporulation.

For more information, go to Newcastle iGEM 2009 [http://2009.igem.org/Team:Newcastle/Metals Metal Sequester] and [http://2009.igem.org/Team:Newcastle/Project/Overview Overview] pages.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 1234
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1046

References

Cretì, P., F. Trinchella, et al. "Heavy metal bioaccumulation and metallothionein content in tissues of the sea bream Sparus aurata from three different fish farming systems." Environmental Monitoring and Assessment.
Morby, A. P., J. S. Turner, et al. (1993). SmtB is a metal-dependent repressor of the cyanobacterial metallothionein gene smtA: identification of a Zn inhibited DNA-protein complex. 21: 921-925.
Waldron, K. J. and N. J. Robinson (2009). "How do bacterial cells ensure that metalloproteins get the correct metal?" Nat Rev Micro 7(1): 25-35.

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 <partinfo>BBa_K174008 short</partinfo>
-SmtA metallothionein protein from ''E. coli'' can bind to heavy metals [1,2,3]. They have a tendency to bind to cationic metal ions such as cadmium, copper, arsenic, mercury, silver. By fusing CotC spore coat protein from ''Bacillus subtilis'', it can be localized to the spore coat, hence trap the metals into bacterial spores. It is also fused with Gfp to see the spores using a microscope.
+SmtA metallothionein protein from ''E. coli'' can bind to heavy metals [1,2,3]. Metallothioneins have a tendency to bind to cationic metal ions such as cadmium, copper, arsenic, mercury, silver.
-We designed this device to sequester cadmium into bacterial spores. It is controlled with ''sigK'' promoter which becomes active when sporulation conditions become active. Hence metals are soaked up when the cells are ready for sporulation. We intended to knock out germination genes and locate SmtA proteins to the spores making the cadmium bio-unavailable.
+By translationally fusing CotC spore coat protein from ''Bacillus subtilis'', SmtA can be localized to the spore coat, hence trapping heavy metals into bacterial spores. SmtA is also fused with Gfp to facilitate the visualisation of spores using a fluorescent microscope.
+We designed this device to sequester cadmium into ''B. subtilis'' spores. The device is expressed using a ''sigK'' type promoter which becomes active under sporulation conditions. Hence metals are soaked up when the cells are undergoing sporulation.
 For more information, go to Newcastle iGEM 2009 [http://2009.igem.org/Team:Newcastle/Metals Metal Sequester] and [http://2009.igem.org/Team:Newcastle/Project/Overview Overview] pages.