Escherichia coli cells which contain a functional Kup (formerly TrkD) system took up Cs' with a moderate rate and affinity. Kup is a separate K+ uptake system with relatively little discrimination in the transport of the cations K+, Rb+, and Cs'. Regardless of the presence or absence of Kup, K+-replete cells took up Cs'primarily by a very low affinity mode, proportional to the ratio of the Cs' and K+ concentrations in the medium. In our project, trkD is used to collect Cs-137 when recN is activated by radiation and start the transcription of trkD,in order to absorb this kind of radioactive source like Cs-137.
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== Expression in ''E. Coli'' ==
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Before we started our functional test of TrkD, we have utilized western blot to determine whether there are enough soluble proteins in E. Coli. With 6×His tag on pET-32a, the result shows that TrkD has more expression with induction of IPTG.
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Note: Because the promoter is Plac here, the expression has a background, which means control also expresses some TrkD without induction of IPTG.
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[[Image:TrkD.jpg]]
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== Detailed Paramers in Result Report ==
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One of the test reports is below. If you need more, please contact me.
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Before test, our samples have been treated with below methods.
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1) Induction of 100mM IPTG
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2) 37℃ for 4h, 200r.
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3) OD: 0.6~1.0
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4）400nmol cesium orginally
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5) use ddH2O to clean the bacterial over 3 times.
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[[Image:Cs Report.jpg]]
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Revision as of 08:31, 9 October 2011
trkD, a functional Kup (formerly TrkD) system took up Cs+ with a moderate rate and affinity
