Difference between revisions of "Part:BBa K606040:Experience"
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
  
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===Applications of BBa_K606040===
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The Hyperspank promoter was designed to be compatible for B. subtilis. It is shorter than the classic one, but is also recognised by E. coli polymerases.
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The Hyperspank promoter was designed to be compatible for B. subtilis. It is shorter thant the classic one, but is also recognised by E. coli polymerases.
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We cloned it first behind the amber tRNA that would induce a GFP amber on an other plasmid. Then to characterise it we cloned it behind an RFP, into TURBO cells that are LacIq to allow the promoter to be the less leacky possible.
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We characterised it with an RFP expression system (PHs-RBS-RFP), into TURBO cells that express LacI to allow the promoter to be the less leacky possible. This system is inducible by IPTG.
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<br><br>
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Without IPTG<br><br><br>
 
{| border="1" class="wikitable" style="text-align: center;"
 
{| border="1" class="wikitable" style="text-align: center;"
 
|E.coli containing Hyperspank+RBS+RFP - IPTG at 37°C
 
|E.coli containing Hyperspank+RBS+RFP - IPTG at 37°C
 
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|[[File:minusfluo1.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (rfp image)]]
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|[[Image:minusfluo1.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (rfp image)]]
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|[[File:minustrans1.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (trans image)]]
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|[[Image:minustrans1.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (trans image)]]
 
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|[[File:minusfluo2.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (rfp image)]]
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|[[Image:minusfluo2.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (rfp image)]]
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|[[File:minustrans2.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (trans image)]]
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|[[Image:minustrans2.jpg|450px|thumb|center|-IPTG E.coli at 37°C  (trans image)]]
 
|}
 
|}
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<br>
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With IPTG : <br><br>
 
{| border="1" class="wikitable" style="text-align: center;"
 
{| border="1" class="wikitable" style="text-align: center;"
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|E.coli containing Hyperspank+RBS+RFP - IPTG at 37°C
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|E.coli containing Hyperspank+RBS+RFP + IPTG at 37°C
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|[[File:hovigfluo1.jpg|450px|thumb|center|+IPTG E.coli at 37°C  (rfp image)]]
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|[[File:hovigtrans1.jpg|450px|thumb|center|+IPTG E.coli at 37°C (trans image)]]
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|[[File:hovigfluo2.jpg|450px|thumb|center|+IPTG E.coli at 37°C  (rfp image)]]
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|[[Image:hovigfluo1.jpg|450px|thumb|center|+IPTG E.coli at 37°C  (rfp image)]]
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|[[File:hovigtrans2.jpg|450px|thumb|center|+IPTG E.coli at 37°C  (trans image)]]
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|[[Image:hovigtrans1.jpg|450px|thumb|center|+IPTG E.coli at 37°C  (trans image)]]
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|-
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|[[Image:hovigfluo2.jpg|450px|thumb|center|+IPTG E.coli at 37°C  (rfp image)]]
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|[[Image:hovigtrans2.jpg|450px|thumb|center|+IPTG E.coli at 37°C  (trans image)]]
 
|}
 
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<partinfo>BBa_K143064 AddReview 5</partinfo>
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This part was used as a part of the Bee subtilis, iGEM METU 2013 project. We simulated the production of RFP under two different IPTG concentrations to observe its basal and maximum transcription rates.
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<!-- DON'T DELETE --><partinfo>BBa_K606040 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K606040 EndReviews</partinfo>

Latest revision as of 02:39, 5 October 2013


The Hyperspank promoter was designed to be compatible for B. subtilis. It is shorter than the classic one, but is also recognised by E. coli polymerases.

We characterised it with an RFP expression system (PHs-RBS-RFP), into TURBO cells that express LacI to allow the promoter to be the less leacky possible. This system is inducible by IPTG.



Without IPTG


E.coli containing Hyperspank+RBS+RFP - IPTG at 37°C
-IPTG E.coli at 37°C (rfp image)
-IPTG E.coli at 37°C (trans image)
-IPTG E.coli at 37°C (rfp image)
-IPTG E.coli at 37°C (trans image)


With IPTG :

E.coli containing Hyperspank+RBS+RFP + IPTG at 37°C
+IPTG E.coli at 37°C (rfp image)
+IPTG E.coli at 37°C (trans image)
+IPTG E.coli at 37°C (rfp image)
+IPTG E.coli at 37°C (trans image)

User Reviews

UNIQc51ab6529bb1187b-partinfo-00000000-QINU

•••••

This part was used as a part of the Bee subtilis, iGEM METU 2013 project. We simulated the production of RFP under two different IPTG concentrations to observe its basal and maximum transcription rates.

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UNIQc51ab6529bb1187b-partinfo-00000002-QINU