Difference between revisions of "Part:BBa K1230000:Experience"
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− | <partinfo>BBa_K1230000 AddReview | + | <partinfo>BBa_K1230000 AddReview 1</partinfo> |
− | <I> | + | <I>iGEM_Nagahama_2016</I> |
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− | We improved the characterization of a previously existing BioBrick Part | + | We improved the characterization of a previously existing BioBrick Part BBa_K1230000 and (https://parts.igem.org/Part:BBa_K1230000) the characterization of a previously existing BioBrick marAgene as BBa_K1653006 . In existing part's information of marA, it gives E. coli resistance against kanamycin and give E-Coli resistance against geraniol as one of the terpene and decrease its intracellular concentration. In this year, we confirmed that overexpressing of marA gives E. coli resistance against farnesol as one of the terpene. This information is very beneficial for other iGEMers to production of toxic organic substances that are produced using bacteria. We introduce an activator gene of AcrAB-TolC efflux pump (MarA) to release the farnesol from the cells and increase the content in the media that shows increase these flavors in the "Flavolator". In our study, we confirmed that overexpressing of marA gives host E. coli high resistance against farnesol. |
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====Resistance==== | ====Resistance==== | ||
− | [[File: | + | [[File:E.colimarA result .jpg|800px|thumb|center|Fig.7:Colony formation of ''E.coli'' JM109 engineered with marA on farnesol overlaid plates. |
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+ | ''E.coli'' JM109 and ''E.coli'' JM109 (marA) were dropped on LBGMg agar plates in serial ten-fold dilutions (10⁻¹~〖10〗^(-6)), overlaid with 30 % (v/v) farnesol hexane solution (farnesol solution) and incubated at 30°C for 24 h. This figure shows that ''E.coli'' JM109 (marA) cells that overexpress the marA product better than the control ''E.coli'' JM109 wild type cells survived plates overloyed by 30 % farnesol solution. | ||
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Revision as of 18:01, 19 October 2016
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K1230000
User Reviews
UNIQ2c2386a8c2cb2bec-partinfo-00000000-QINU
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We were able to characterise this part, since we found a measurable phenotype (kanamycin resistance).
A test was performed with three different antibiotics and different IPTG concentrations. The part seems to make the host sensible to Tetracyclin (maybe because it's on Cm plasmid, and both antibiotics act on similar ways), it seems not to have an effect on Ampicilin and it shows a relevant effect with Kanamycin. Which is further explored in the next figure.
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UNIQ2c2386a8c2cb2bec-partinfo-00000002-QINU
Nagahama_2015
UNIQ2c2386a8c2cb2bec-partinfo-00000003-QINU
•••
iGEM_Nagahama_2015 |
The activator of AcrAB-TolC multidrug efflux pump exports some terpenes
Export geraniol
ResistanceNagahama_2016UNIQ2c2386a8c2cb2bec-partinfo-00000005-QINU
UNIQ2c2386a8c2cb2bec-partinfo-00000007-QINU |