Difference between revisions of "Part:BBa K1653006"
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+ | ====Export geraniol==== | ||
+ | [[File:GC conc fig.png |650px|thumb|center|Fig. 1: Intracellular geraniol concentrations of ''E. coli'' JM109 (WT) and its overexpressing of ''marA'' strain, ''E. coli'' JM109 (''marA''). | ||
+ | <br> | ||
+ | In this figure, intracellular content of geraniol was less in the strain ''E. coli'' JM109 (''marA'') than the strain ''E. coli'' JM109 (WT). The concentrations of intracellular geraniol from ''E. coli'' JM109 (''marA'') was 42.9 μg/ml, which was 40% lower than that from of ''E. coli'' JM109 (WT), 72.2 μg/ml. This figure is suggesting that internalized geraniol could be more efficiently exported through AcrAB-TolC efflux pump following the presumed activation of this gene by introducing the activator ''marA'' gene.]] | ||
+ | <br> | ||
+ | The following graphs show that overexpressing marA gives resistance against geraniol and decrease intracelluler geraniol concentration. | ||
+ | <br>'''[https://parts.igem.org/Part:BBa_K1653020 ''marA'' device BBa_K16530020]''' | ||
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+ | ====Resistance==== | ||
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+ | [[File:MarA plate assay yoshiharu did.png|750px|thumb|center|Fig. 2: Colony formation efficiencies of E. coli JM109 engineered with marA on geraniol overlaid plates. | ||
+ | E. coli JM109 and E. coli JM109 (marA) were spotted on LBGMg agar plates in serial ten-fold dilutions (10⁻¹~10⁻⁵), overlaid with geraniol solutions, and incubated at 30°C for 24 h. This figure shows that E. coli JM109 (marA) cells that overexpress the marA product is more survived on geraniol overlay plates than the counterpart control E. coli JM109 wild type cells. | ||
+ | ]] | ||
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+ | [[File:MarA plate assay nishikawa last.png|650px|thumb|center|Fig. 3: Comparison of colony numbers after addition of geraniol solution. Time interval for treatment was set every 1 hour from 1 hour to 4 hours. A: ''E. coli'' JM109 (WT) + hexane; B: ''E. coli'' JM109 (''marA'') + hexane; C: ''E. coli'' JM109 (WT) + geraniol; D: ''E. coli'' JM109 (''marA'') + geraniol. | ||
+ | As shown in Figs. 2 A and B, treatment with hexane of ''E. coli'' JM109 (WT) and of ''E. coli'' JM109 (''marA'') showed similar colony numbers during these treatment intervals to those of time zero. This result suggests that hexane at this concentration and duration of time for 4hours did not affect both cell growth. In contrast, treatment with geraniol of ''E. coli'' JM109 (WT) and of ''E. coli'' JM109 (''marA'') showed toxicities to both strains (Figs. 3 B, C and D). If we watch the colony numbers carefully, ''E. coli'' JM109 (''marA'') had more than ''E. coli'' JM109 (WT) during these treatment intervals ((Figs. 3 C and D). These results demonstrate that toxicity of the geraniol was less to the strain ''E. coli'' JM109 (''marA'') than the strain ''E. coli'' JM109 (WT). | ||
+ | ]] | ||
Revision as of 09:09, 17 September 2015
MarA
The activator of AcrAB-TolC multidrug efflux pump exports some terpenes
We improved the characterization of a previously existing BioBrick Part BBa_K1230000 and submitted this improved BioBrick marA device as BBa_K1653006 to iGEM Registry.
In exsisting part's information of marA, it gives E. coli resistance against kanamycin only.
In this year, we confilmed that overepressing of marA gives E. coli resistance against geraniol as one of the terpene and decrease its intracellular concentration. This information is very beneficial for other iGEMers to production of toxic organic substances that are produced using bacteria.
[Results are here]
Export geraniol
![](/wiki/images/c/c1/GC_conc_fig.png)
In this figure, intracellular content of geraniol was less in the strain E. coli JM109 (marA) than the strain E. coli JM109 (WT). The concentrations of intracellular geraniol from E. coli JM109 (marA) was 42.9 μg/ml, which was 40% lower than that from of E. coli JM109 (WT), 72.2 μg/ml. This figure is suggesting that internalized geraniol could be more efficiently exported through AcrAB-TolC efflux pump following the presumed activation of this gene by introducing the activator marA gene.
The following graphs show that overexpressing marA gives resistance against geraniol and decrease intracelluler geraniol concentration.
marA device BBa_K16530020
Resistance
![](/wiki/images/4/41/MarA_plate_assay_yoshiharu_did.png)
![](/wiki/images/4/4d/MarA_plate_assay_nishikawa_last.png)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 148
- 1000COMPATIBLE WITH RFC[1000]