Difference between revisions of "Part:BBa K2483000"

Revision as of 11:00, 7 September 2017

IAA producing enzymes fused to RNA-binding proteins under control of Pveg2

This part is an upgrade to the part BBa_K515100 of iGEM Imperial College 2012 producing Indoleacetic acid with the enzymes IaaM (Indoleacetic Acid tryptophan monooxygenase) and IaaH (Indoleacetamide Hydrolase). Here we just fused the IAA enzymes to the RNA-binding proteins (RBPs) MS2 and PP7, respectively.

We used this part to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by putting them in close proximity to each other (metabolic channelling). This was done with a dCas9 (BBa_K2483002) scaffold where the RNA-binding proteins bind to aptamer sequences added to the 3'-end of a sgRNA (BBa_K2483003) and the sgRNA binds to target cassettes where several of the sgRNA recognition sequences are located behind each other (BBa_K2483005 and BBa_K2483006).

For the complete part with sgRNA, dCas9 and the IAA-enzymes, see BBa_K2483004.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 547
Illegal BamHI site found at 1492
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 254
Illegal NgoMIV site found at 3198
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2483000 short</partinfo>
-This is a fusion protein of IaaM  (Indoleacetic Acid tryptophan monooxygenase (IaaM) BBa_K515000) catalysing the oxidative carboxylation of L-tryptophan to indole-3-acetamide and MCP a RNA-binding protein from the virus MS2. Additionally, we've included the pVeg2 promoter and rbs (BBa_K5151010) so this part is essentially the part K515100 of Team Imperial College 2011 without the IaaH and with MCP fused to IaaM.
+This part is an upgrade to the part BBa_K515100 of iGEM Imperial College 2012 producing Indoleacetic acid with the enzymes IaaM (Indoleacetic Acid tryptophan monooxygenase) and IaaH (Indoleacetamide Hydrolase). Here we just fused the IAA enzymes to the RNA-binding proteins (RBPs) MS2 and PP7, respectively.
-We used this part (in conjunction with BBa_K2483001) to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by puttign them in close proximity to each other (metabolic channelling). This was done with a dCas9 scaffold where the RNA-binding proteins bind to aptamer sequences added to the 3'-end of sgRNA and the sgRNA binds to target cassettes where several of the sgRNA recognition sequences are located behind each other (BBa_K2483005 and BBa_K2483006).
-There is another part with IaaH fused to PP7, another RNA-binding protein called BBa_K2483001. For the complete part with sgRNA, dCas9 and the IAA-enzymes, see BBa_K2483004.
+We used this part to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by putting them in close proximity to each other (metabolic channelling). This was done with a dCas9 (BBa_K2483002) scaffold where the RNA-binding proteins bind to aptamer sequences added to the 3'-end of a sgRNA (BBa_K2483003) and the sgRNA binds to target cassettes where several of the sgRNA recognition sequences are located behind each other (BBa_K2483005 and BBa_K2483006).
+For the complete part with sgRNA, dCas9 and the IAA-enzymes, see BBa_K2483004.
 <!-- Add more about the biology of this part here