IGEM:IMPERIAL/2006/project/Oscillator/project browser/Test Sensing Predator Construct/Implementation: Difference between revisions

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|-
|-
!height="25pt" width="80pt"|Actual Part
!height="25pt" width="80pt"|Actual Part
!colspan="4"| Logo of the Part
!colspan="4"| [[Image:J37016 logo.png|300px]]
|-
|-
!height="25pt" width="80pt"|Sub Parts
!height="25pt" width="80pt"|Sub Parts
! [[IGEM:IMPERIAL/2006/project/Oscillator/project browser/intermediate_part| intermediate_part]]
! <bbpart>R0062</bbpart>
! [[IGEM:IMPERIAL/2006/project/Oscillator/project browser/intermediate_part| intermediate_part]]
! <bbpart>B0034</bbpart>
! [[IGEM:IMPERIAL/2006/project/Oscillator/project browser/intermediate_part| intermediate_part]]
! <bbpart>C0062</bbpart>
! [[IGEM:IMPERIAL/2006/project/Oscillator/project browser/intermediate_part| intermediate_part]]
! <bbpart>I13504</bbpart>
|}
|}
<div class="tabs-blue">
<div class="tabs-blue">
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</ul>
</ul>
</div>
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<br style="clear:both">
<br style="clear:both">


These two parts are very similar and have the same inputs and outputs, therefore they can be analysed in exactly the same way
==Ligation strategy==


<br>
<u><font size="2">'''Ligating the Polycystronic Predator 'Sensing' Test Construct:'''</font></u>
*<big>'''[http://parts.mit.edu/registry/index.php/Part:BBa_J37016 J37016]'''</big>
__NOTOC__
<br>
{| border="1" cellpadding="0" width="100%"
[[image:J37016.JPG]]
|-
!style="background:#E6B3E0" height="25pt" width="80pt" |For !! style="background:#E6B3E0" |Stage 1 !! style="background:#E6B3E0" |Stage 2 !! style="background:#E6B3E0" |Stage 3
|-
!style="background:#D9F2ED"|<center><bbpart>J37033</bbpart>=<br>(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)</center> !!style="background:#CCEBAD" | <center>[[Image:B0034 logo.png]][[Image:Plus sign.png|20px]][[Image:C0062 logo.png]]</center>
|-
!style="background:#D9F2ED"|<center><bbpart>J37019</bbpart>=<br>(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)</center>|| !!style="background:#CCEBAD" | <center>[[Image:R0062 logo.png]][[Image:Plus sign.png|20px]][[Image:B0034 logo.png]][[Image:C0062 logo.png]]</center>
|-
!style="background:#D9F2ED"|<center><bbpart>J37016</bbpart>=<br>(<bbpart>J37019</bbpart>+<bbpart>J37025</bbpart>)</center>|| || !!style="background:#CCEBAD" | <center>[[Image:J37016 logo.png]]</center>
|}


This construct is used to characterise the pLuxR transfer function, and noteably its effect on YFP expression, which would be equivalent to AiiA expression in the full system with one promoter.
<u><font size="2">'''Ligating the Monocystronic Predator 'Sensing' Test Construct:'''</font></u>
<br>
__NOTOC__
<br>
{| border="1" cellpadding="0" width="100%"
*[http://parts.mit.edu/registry/index.php/Part:BBa_R0062 R0062] - 9G Plate 1
|-
*[http://parts.mit.edu/registry/index.php/Part:BBa_B0034 B0034] - 3O Plate 1
!style="background:#E6B3E0" height="25pt" width="80pt" |For !! style="background:#E6B3E0" |Stage 1 !! style="background:#E6B3E0" |Stage 2 !! style="background:#E6B3E0" |Stage 3 !! style="background:#E6B3E0" |Stage 4
*[http://parts.mit.edu/registry/index.php/Part:BBa_C0062 C0062] - 7A Plate 1
|-
*[http://parts.mit.edu/registry/index.php/Part:BBa_I13504 I13504] - 12D Plate 1
!style="background:#D9F2ED"|<center><bbpart>J37033</bbpart>=<br>(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)</center> !! style="background:#CCEBAD"| <center>[[Image:B0034 logo.png]][[Image:Plus sign.png|20px]][[Image:C0062 logo.png]]</center>
|-
!style="background:#D9F2ED"|<center><bbpart>J37019</bbpart>=<br>(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)</center>|| !!style="background:#CCEBAD" |<center>[[Image:R0062 logo.png]][[Image:Plus sign.png|20px]][[Image:B0034 logo.png]][[Image:C0062 logo.png]]</center>
|-
!style="background:#D9F2ED"|<center><bbpart>J37031</bbpart>=<br>(<bbpart>J37019</bbpart>+<bbpart>B0014</bbpart>)</center>|| || !!style="background:#CCEBAD" | [[Image:J37031 logo.png]]
|-
!style="background:#D9F2ED"|<center><bbpart>J37032</bbpart>=<br>(<bbpart>R0062</bbpart>+<bbpart>I13504</bbpart>)</center> !!style="background:#CCEBAD" | [[Image:R0062 logo.png]][[Image:Plus sign.png|20px]][[Image:I13504 logo.png]]
|-
!style="background:#D9F2ED"|<center><bbpart>J37020</bbpart>=<br>(<bbpart>J37031</bbpart>+<bbpart>J37032</bbpart>)</center>|| || || !!style="background:#CCEBAD" | [[Image:J37020 logo.png]]
|}


*More information on ligations can be found on the [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/Ligation_Page Ligation Page].


<br>
==Quality control==
*<big>'''[http://parts.mit.edu/registry/index.php/Part:BBa_J37020  J37020]'''</big>
<br>
[[image:J37020.JPG]]


This construct is used to characterise the pLuxR transfer function, and noteably its effect on YFP expression, which would be equivalent to AiiA expression in the full system with two promoters.
__NOTOC__
{| border="1" cellpadding="0" width="100%"
|-
!height="25pt" width="80pt"| Part !! Logo !! Built !! Sequenced !! Primers Used for Sequencing
|-
| <center><bbpart>J37016</bbpart></center> || [[Image:J37016 logo.png]] || Yes || [http://openwetware.org/images/4/4d/J37016_sequencing.doc Yes] || [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/parts/Primers Primers]
|-
| <center><bbpart>J37019</bbpart></center> || [[Image:J37019 part.png]] || Yes || [http://openwetware.org/images/e/e3/J37019_sequencing.doc Yes] || [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/parts/Primers Primers]
|-
| <center><bbpart>J37020</bbpart></center> || [[Image:J37020 logo.png]] || No || N/A || N/A
|-
| <center><bbpart>J37031</bbpart></center> || [[Image:J37031 logo.png]] || Yes || [http://openwetware.org/images/a/ac/J37020sequencing.doc Yes] || [http://openwetware.org/wiki/IGEM:IMPERIAL/2006/project/parts/Primers Primers]
|-
| <center><bbpart>J37032</bbpart></center> || [[Image:J37032 logo.png]] || Yes || No || N/A
|-
| <center><bbpart>J37033</bbpart></center> || [[Image:J37033 logo.png]] || Yes || No || N/A
|}


The input is HSL and the outputs are LuxR and GFP. GFP will be mesured
==WetLab work entries==


*[http://parts.mit.edu/registry/index.php/Part:J37018 J37018] - To be built!
*The polycystronic predator sensing part (J37016) has been successfully built, and tested.  
*[http://parts.mit.edu/registry/index.php/Part:I13504 I13504] - 12D Plate 1
*The monocystronic predator sensing part (J37020) was not built.

Latest revision as of 07:47, 30 October 2006

Super Parts Predator Construct
Actual Part
Sub Parts <bbpart>R0062</bbpart> <bbpart>B0034</bbpart> <bbpart>C0062</bbpart> <bbpart>I13504</bbpart>


Ligation strategy

Ligating the Polycystronic Predator 'Sensing' Test Construct:

For Stage 1 Stage 2 Stage 3
<bbpart>J37033</bbpart>=
(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)
<bbpart>J37019</bbpart>=
(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)
<bbpart>J37016</bbpart>=
(<bbpart>J37019</bbpart>+<bbpart>J37025</bbpart>)

Ligating the Monocystronic Predator 'Sensing' Test Construct:

For Stage 1 Stage 2 Stage 3 Stage 4
<bbpart>J37033</bbpart>=
(<bbpart>B0034</bbpart>+<bbpart>C0062</bbpart>)
<bbpart>J37019</bbpart>=
(<bbpart>R0062</bbpart>+<bbpart>J37033</bbpart>)
<bbpart>J37031</bbpart>=
(<bbpart>J37019</bbpart>+<bbpart>B0014</bbpart>)
<bbpart>J37032</bbpart>=
(<bbpart>R0062</bbpart>+<bbpart>I13504</bbpart>)
<bbpart>J37020</bbpart>=
(<bbpart>J37031</bbpart>+<bbpart>J37032</bbpart>)

Quality control

Part Logo Built Sequenced Primers Used for Sequencing
<bbpart>J37016</bbpart>
 Yes Yes Primers
<bbpart>J37019</bbpart>
 Yes Yes Primers
<bbpart>J37020</bbpart>
 No N/A N/A
<bbpart>J37031</bbpart>
 Yes Yes Primers
<bbpart>J37032</bbpart>
 Yes No N/A
<bbpart>J37033</bbpart>
 Yes No N/A

WetLab work entries

  • The polycystronic predator sensing part (J37016) has been successfully built, and tested.
  • The monocystronic predator sensing part (J37020) was not built.
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