Difference between revisions of "Part:BBa K911002:Experience"

(→‎Applications of BBa_K911002)
(→‎Applications of BBa_K911002)
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[[Image:MgConstruct1.jpg|800px|center]]
 
[[Image:MgConstruct1.jpg|800px|center]]
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Unfortunately, we were unable to get this part to insert into our vector, we were unable to characterize it.
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Because our experiments without the eight codon substitution [[Part:BBa_K911001:Experience|failed]], future teams may wish to repeat our attempts at testing this part with the eight codon substitution. We do not recommend using lacI as part of your reporter, as this protein require its N-terminal for its function. Instead, an appropriate fluorescent protein may be used, though take care that any responsiveness on the part of the magnesium will result in a decrease in fluorescence. Alternatively, the original paper successfully used lacZ as a reporter, as shown.
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[[Image:MgPaperAssay.jpg|500px|center|thumb|Successful test of the magnesium riboswitch by the Dann et al. team. Bars represent β-galactosidase activity, as determined by a Miller assay at hourly intervals (at 30 μM and 10 mM concentrations of Mg2+ - low and high respectively). Lines represent bacterial growth, quantified by the left hand y-axis. From Dann et al. Cell (2007)]]
  
 
===User Reviews===
 
===User Reviews===

Revision as of 23:02, 25 September 2012

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_K911002

We attepted to insert this part into the following assembly, using Gibson assembly:

MgConstruct1.jpg

Unfortunately, we were unable to get this part to insert into our vector, we were unable to characterize it.

Because our experiments without the eight codon substitution failed, future teams may wish to repeat our attempts at testing this part with the eight codon substitution. We do not recommend using lacI as part of your reporter, as this protein require its N-terminal for its function. Instead, an appropriate fluorescent protein may be used, though take care that any responsiveness on the part of the magnesium will result in a decrease in fluorescence. Alternatively, the original paper successfully used lacZ as a reporter, as shown.

Successful test of the magnesium riboswitch by the Dann et al. team. Bars represent β-galactosidase activity, as determined by a Miller assay at hourly intervals (at 30 μM and 10 mM concentrations of Mg2+ - low and high respectively). Lines represent bacterial growth, quantified by the left hand y-axis. From Dann et al. Cell (2007)

User Reviews

UNIQ37bbd49db0be8c4a-partinfo-00000000-QINU UNIQ37bbd49db0be8c4a-partinfo-00000001-QINU