Difference between revisions of "Part:BBa K2075020"

Latest revision as of 18:59, 18 October 2016

TAL-effector Ax7L-DS

This part synthesize a transcription activator like (TAL) effector protein. They were found in Xanthomonas bacteria. These bacteria are plant pathogenes and infects plants via a secretion typ III system. In the plant cells these proteins binds promoter sites. This leads to a transcriptional activation of the genes behind the promoter. The proteins contains tandem repeats up to 34.5. Each repeat binds one nucleotide of the target sequenz. The 12th and 13th amino acid of each repeat leads to the specific binding of the DNA sequence. We circularised the TAL-effectors with the help of Inteins at the N- and C- Terminus of each TAL-effector. The 12th and 13th amino acid of each of the repeats from our TAL effector Ax7L-DS are: NI NN HD NI HD NG NI NG NI NG NI NI NI HD HD HD HD HD They bind the DNA sequence: A G/A C A C T A T A T A A A C C C C C

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 3202
Illegal BamHI site found at 2608
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 2547
Illegal NgoMIV site found at 3142
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 2622

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2075020 short</partinfo>
-Designing cyclic TALEs allows a regulation of those proteins, because of topological problems. A TALE is always winding itself around the DNA to bind. If the protein is cyclic, this is no longer possible and the TALE-bond is inhibited. This could also be used for appliances concerning drug delivery. If the cyclic bond is irreversible and a protease can cut the protein, the TALE regains full transcriptional activity (Lonzari&#263;, 2016). To prove this statement, we inserted a TEV cleavage site from the tobacco each virus into the vector which enables induced linearization of the protein after expression with ProTEV Plus protease (Promega).
+This part synthesize a transcription activator like (TAL) effector protein. They were found in Xanthomonas bacteria. These bacteria are plant pathogenes and infects plants via a secretion typ III system. In the plant cells these proteins binds promoter sites. This leads to a transcriptional activation of the genes behind the promoter. The proteins contains tandem repeats up to 34.5. Each repeat binds one nucleotide of the target sequenz. The 12th and 13th amino acid of each repeat leads to the specific binding of the DNA sequence.
+We circularised the TAL-effectors with the help of Inteins at the N- and C- Terminus of each TAL-effector.
-This composite part includes an Intein and linker, the TAL-effector Ax7L-DS, a TEV Site and a Strep XT Tag. They are translated all together.
+The 12th and 13th amino acid of each of the repeats from our TAL effector Ax7L-DS are: NI NN HD NI HD NG NI NG NI NG NI NI NI HD HD HD HD HD They bind the DNA sequence: A G/A C A C T A T A T A A A C C C C C