Difference between revisions of "Part:BBa K1416001"

Latest revision as of 01:22, 17 October 2014

The tRNA synthetase/tRNA needed for incorporating 3-iodo-L-tyrosine (IodoY) at a UAG codon

The part includes the gene coding for the Methanocaldococcus jannaschii synthetase that charges the orthogonal tRNA (also in the part) with the amino acid 3-iodotyrosine along with the proper promoters and terminator. This part encodes for the incorporation of tyrosine during translation at AUG amber stop codons in a cell with Release Factor 1 (RF1) knocked out such as "Amberless" E. coli or "RF0".

This part was used by the UT Austin 2014 iGEM team in their [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project]. In this work they demonstrated that this part was one of the best ncAA tRNA synthetase/tRNA pairs

In the control cultures, IodoY-pFRYC, the level of GFP expression relative to RFP expression is set as a standard. In the test cultures, IodoY-pFRY, the level of GFP expression relative to RFP expression indicates how well the part function. In the absence of ncAA, GFP expression is about 20% of RFP expression, a much lower value than in the control conditions. In the presence of ncAA, the amount of GFP expression increases back to roughly 100% of RFP expression. These results indicate that part BBa_K1416001 has good fidelity, and is very efficient. For more information and details, please visit [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project]

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 1153
Illegal BamHI site found at 1159
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1185
Illegal NgoMIV site found at 1645
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 316
Illegal SapI.rc site found at 1109

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1416001 short</partinfo>
-The part includes the gene coding for the <i>Methanocaldococcus jannaschii</i> synthetase that charges the orthogonal tRNA (also in the part) with the amino acid tyrosine along with the proper promoters and terminator. This part encodes for the incorporation of tyrosine during translation at AUG amber stop codons in a cell with Release Factor 1 (RF1) knocked out such as "Amberless" E. coli or "RF0".
+The part includes the gene coding for the <i>Methanocaldococcus jannaschii</i> synthetase that charges the orthogonal tRNA (also in the part) with the amino acid 3-iodotyrosine along with the proper promoters and terminator. This part encodes for the incorporation of tyrosine during translation at AUG amber stop codons in a cell with Release Factor 1 (RF1) knocked out such as "Amberless" E. coli or "RF0".
+This part was used by the UT Austin 2014 iGEM team in their [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project]. In this work they demonstrated that this part was one of the best ncAA tRNA synthetase/tRNA pairs
+[[File:IodoY-UT_Austin_2014.png|thumb|500px|center| In the control cultures, IodoY-pFRYC, the level of GFP expression relative to RFP expression is set as a standard.  In the test cultures, IodoY-pFRY, the level of GFP expression relative to RFP expression indicates how well the part function.  In the absence of ncAA, GFP expression is about 20% of RFP expression, a much lower value than in the control conditions.  In the presence of ncAA, the amount of GFP expression increases back to roughly 100% of RFP expression.  These results indicate that part BBa_K1416001 has good fidelity, and is very efficient.  For more information and details, please visit [http://2014.igem.org/Team:Austin_Texas/kit#Results_and_Data ncAA kit project]]]
 <!-- Add more about the biology of this part here
 ===Usage and Biology===