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Applications of BBa_K1230000
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 organic substance that have toxicity using bacteria.
Resistance
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 (10â»Â¹ï½ž10â»âµ), 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.
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).
Fig 4. Colony numbers of Fig3 A~D
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UNIQ534fd857cff29970-partinfo-00000000-QINU
UNIQ534fd857cff29970-partinfo-00000001-QINU
