Difference between revisions of "Part:BBa K902017"

Latest revision as of 09:34, 29 October 2012

MgtAp-mgtArb-GFP(LVA)

This part was built to characterize the control element of the magnesium killswitch with mgtAp-mgtArb. GFP is used as a reporter to indicate how well the system responds to different concentrations of magnesium compared to controls. For the protocol and experimental design please click here. For characterization data please click here. This part shows that with lower concentrations of magnesium it is activated and with higher concentrations of magnesium it is repressed.

Figure 1: The blue line indicating the response to magnesium shows a dramatic decrease at approximately 1 mM concentration of magnesium. the system completely shuts off at 1mM magnesium.

It is important to note that these constructs do not contain the signal transduction components PhoP and PhoQ and therefore the promoter riboswitch system appears to be leaky, i.e it does not completely shut off when there is 10mM magnesium.

Usage and Biology

The mgt system is used in the control and metabolism of magnesium in E. coli. It is important in regulating the transport of these ions into and out of the cell to ensure homeostasis is maintained. It has been previously characterized that the mgtA transport system contains an upstream promoter and riboswitch sensative to magnesium. We have cloned and biobricked both of these components and shown that they demonstrate the ability

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 974

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 <partinfo>BBa_K902017 short</partinfo>
-<html> This part was built to characterize the control element of the magnesium killswitch with mgtAp-mgtArb. GFP is used as a reporter to indicate how well the system responds to different concentrations of magnesium compared to controls. For the protocol and experimental design please click <a href="http://2012.igem.org/Team:Calgary/Notebook/Protocols/mgcircuit">here</a> For characterization data please click <a href="http://2012.igem.org/Team:Calgary/Project/HumanPractices/Killswitch/Regulation">here</a>. This part shows that with lower concentrations of magnesium it is activated and with higher concentrations of magnesium it is repressed. </html>[[Image:Magmesium graph.png|thumb|center|500px|Figure 1: The blue line indicating the response to magnesium shows a dramatic decrease at approximately 1 mM concentration of magnesium. the system completely shuts off at 1mM magnesium.]]
+<html> This part was built to characterize the control element of the magnesium killswitch with mgtAp-mgtArb. GFP is used as a reporter to indicate how well the system responds to different concentrations of magnesium compared to controls. For the protocol and experimental design please click <a href="http://2012.igem.org/Team:Calgary/Notebook/Protocols/mgcircuit">here</a>. For characterization data please click <a href="http://2012.igem.org/Team:Calgary/Project/HumanPractices/Killswitch/Regulation">here</a>. This part shows that with lower concentrations of magnesium it is activated and with higher concentrations of magnesium it is repressed. </html>[[Image:Magmesium graph.png|thumb|center|500px|Figure 1: The blue line indicating the response to magnesium shows a dramatic decrease at approximately 1 mM concentration of magnesium. the system completely shuts off at 1mM magnesium.]] <html> It is important to note that these constructs do not contain the signal transduction components <a href="https://parts.igem.org/BBa_K902010">PhoP</a> and <a href="https://parts.igem.org/BBa_K902011">PhoQ</a> and therefore the promoter riboswitch system appears to be leaky, i.e it does not completely shut off when there is 10mM magnesium. </html>
 ===Usage and Biology===