Difference between revisions of "Part:BBa K1653020"
Yoshiharu994 (Talk | contribs) |
(→Geraniol resistance) |
||
(12 intermediate revisions by 2 users not shown) | |||
Line 2: | Line 2: | ||
<partinfo>BBa_K1653020 short</partinfo> | <partinfo>BBa_K1653020 short</partinfo> | ||
− | + | <br> | |
− | This device consists of R0011+B0034+K1653006+B0015. | + | This device consists of [https://parts.igem.org/wiki/index.php?title=Part:BBa_R0011 R0011]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_B0034 B0034]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_K1653006 K1653006]+[https://parts.igem.org/wiki/index.php?title=Part:BBa_B0015 B0015]. |
− | + | ||
+ | [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1653006 BBa_K1653006] was improved the characterization of a previously existing BioBrick Part [https://parts.igem.org/Part:BBa_K1230000 BBa_K1230000] | ||
+ | <br> | ||
+ | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
+ | |||
<!-- --> | <!-- --> | ||
Line 17: | Line 21: | ||
<partinfo>BBa_K1653020 parameters</partinfo> | <partinfo>BBa_K1653020 parameters</partinfo> | ||
<!-- --> | <!-- --> | ||
+ | |||
+ | ====Export geraniol==== | ||
+ | <br> | ||
+ | [[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.]] | ||
+ | |||
+ | ====Geraniol resistance==== | ||
+ | |||
+ | [[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. | ||
+ | <br> | ||
+ | ''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 1.0 % (V/V) geraniol hexane solution (geraniol solution), 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 1.0 % geraniol solution overlay plates than the counterpart control ''E. coli'' JM109 wild type cells. | ||
+ | |||
+ | ]] | ||
+ | |||
+ | [[File:MarA plate assay nishikawa last.png|650px|thumb|center|Fig. 3: Comparison of colony numbers after addition of 0.5 %( V/V) geraniol hexane solution (geraniol solution). | ||
+ | <br> | ||
+ | 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) + 0.5 % geraniol solution; D: ''E. coli'' JM109 (''marA'') + 0.5 % geraniol solution. 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). | ||
+ | ]] |
Latest revision as of 14:44, 17 September 2015
marA.dev
This device consists of R0011+B0034+K1653006+B0015.
BBa_K1653006 was improved the characterization of a previously existing BioBrick Part BBa_K1230000
Sequence and Features
Assembly Compatibility:
- 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 229
- 1000COMPATIBLE WITH RFC[1000]
Export geraniol