Difference between revisions of "Part:BBa I742111:Experience"

Latest revision as of 20:13, 3 October 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_I742111

A composite LIMS1 part with a Lac promoter has been characterized. For details, please visit this page.

Team TU Munich 2012 used this part as PCR-template to obtain the coding sequence of (+)-Limonene Synthase. See the page of our BioBrick BBa_K801060.

User Reviews

UNIQ30ea14677133242e-partinfo-00000002-QINU

•••

iGEM Dundee 2011

We found the sequence of this part to be correct as listed and modified it by the addition of a microcompartment targeting domain and an HA epitope tag (as BBa_K562015). This will allow better monitoring of production of the protein, and it was detectable using an anti-HA monoclonal antibody whenexpressed in E. coli. We did not get the chance to test limonene production in a host cell producing this part.

••

iGEM Wisconsin-Madison 2012

The sequence of this part matched the sequence listed. However, when we used this gene for the production of limonene, we were not able to produce any in E. coli. We thought a problem may be rare codons in the sequence, so we codon optimized it for E. coli and sent it in with our parts this year (BBa_K762100). However, neither the regular or codon optimized limonene synthase showed a GC/MS peak near the expected limonene ion. To toubleshoot the lack of limonene, we cloned the limonene synthase part (both codon optimized and non codon optimized) into the translational coupling cassette (BBa_K762000). This part easily determines if a part is being translated or not. According to the data we obtained from this cassete, codon-optimized limonene synthase is being fully translated by E. Coli and the non-codon optimzed version (BBa_I742111) is not being translated at all. See the following page for the TCC data:

https://parts.igem.org/Part:BBa_K762100:Experience#Applications_of_BBa_K762100

In addition to the change in translation, growth curve data shows that the codon optimized version grows better in standard media.

See the following page for growth curve data

http://2012.igem.org/Team:Wisconsin-Madison/lemon

UNIQ30ea14677133242e-partinfo-00000006-QINU

@@ Line 7: / Line 7: @@
 A composite LIMS1 part with a Lac promoter has been characterized. For details, please visit this <html><a href="https://parts.igem.org/Part:BBa_K118025:Experience">page</a>.</html>
+Team TU Munich 2012 used this part as PCR-template to obtain the coding sequence of (+)-Limonene Synthase. See the page of our BioBrick <html><a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_K801060">BBa_K801060</a>.</html>
 ===User Reviews===
@@ Line 19: / Line 21: @@
 |width='60%' valign='top'|
-We found the sequence of this part to be correct as listed and modified it by the addition of a microcompartment targeting domain and an HA epitope tag (as <partinfo>K562018</partinfo>). This will allow better monitoring of production of the protein. We did not get the chance to test arsenic resistance in a host cell producing this part.
+We found the sequence of this part to be correct as listed and modified it by the addition of a microcompartment targeting domain and an HA epitope tag (as <partinfo>K562015</partinfo>). This will allow better monitoring of production of the protein, and it was detectable using an anti-HA monoclonal antibody whenexpressed in <i>E. coli</i>. We did not get the chance to test limonene production in a host cell producing this part.
 |}
+{|width='80%' style='border:1px solid gray'
+|-
+|width='10%'|
+<partinfo>BBa_K190019 AddReview 2</partinfo>
+<I>iGEM Wisconsin-Madison 2012</I>
+|width='60%' valign='top'|
+The sequence of this part matched the sequence listed. However, when we used this gene for the production of limonene, we were not able to produce any in <i>E. coli</i>. We thought a problem may be rare codons in the sequence, so we codon optimized it for <i>E. coli</i> and sent it in with our parts this year (BBa_K762100). However, neither the regular or codon optimized limonene synthase showed a GC/MS peak near the expected limonene ion. To toubleshoot the lack of limonene, we cloned the limonene synthase part (both codon optimized and non codon optimized) into the translational coupling cassette (BBa_K762000). This part easily determines if a part is being translated or not. According to the data we obtained from this cassete, codon-optimized limonene synthase is being fully translated by ''E. Coli'' and the non-codon optimzed version (BBa_I742111) is not being translated at all. See the following page for the TCC data:
+https://parts.igem.org/Part:BBa_K762100:Experience#Applications_of_BBa_K762100
+In addition to the change in translation, growth curve data shows that the codon optimized version grows better in standard media.
+See the following page for growth curve data
+http://2012.igem.org/Team:Wisconsin-Madison/lemon
+|}
 ;
 <!-- End of the user review template -->
 <!-- DON'T DELETE --><partinfo>BBa_K190019 EndReviews</partinfo>