"
Team:Michigan/Notebook
From 2012.igem.org
(Difference between revisions)
| Line 19: | Line 19: | ||
<html> | <html> | ||
| - | <h2> | + | <h2>Increasing Recombinase HbiF and FimE Turnover Rate with LVA Degradation Tag </h2> |
<p>HbiF and LVA degradation tag were ordered from IDT with the following specifications.</p> | <p>HbiF and LVA degradation tag were ordered from IDT with the following specifications.</p> | ||
| Line 37: | Line 37: | ||
<p style="font-family:Courier, monospace;"><span style="background-color:#86C67C;">ACCGGT</span> TAAT <span style="background-color:#C8E0DB">ACTAGT</span> A GCGGCCG <span style="background-color:#A9C9A4">CTGCAG</span> </p> | <p style="font-family:Courier, monospace;"><span style="background-color:#86C67C;">ACCGGT</span> TAAT <span style="background-color:#C8E0DB">ACTAGT</span> A GCGGCCG <span style="background-color:#A9C9A4">CTGCAG</span> </p> | ||
| - | + | <p>In order to properly attach the LVA degradation tag, the prefix of the tag sequence must be complementary to the suffix of Hbif. For this part, the prefix and suffix are reversed in comparison to above: the prefix is RFC25, and the suffix is RFC10, for reasons that are discussed below.</p> | |
Revision as of 00:09, 6 July 2012
Team:Michigan/Notebook/Protocols
| Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
|---|
Increasing Recombinase HbiF and FimE Turnover Rate with LVA Degradation Tag
HbiF and LVA degradation tag were ordered from IDT with the following specifications.
EcoRI
XbaI
SpeI
PstI
AgeI
NgoMIV
HbiF possesses an N-terminus in the (truncated) RFC10 format :
GAATTC GCGGCCGC T TCTAG [ATG]
No extra amino acids are created by this prefix, in comparison to the standard RFC25 prefix, which creates a serine and glycine. The suffix proceeds as follows:
ACCGGT TAAT ACTAGT A GCGGCCG CTGCAG
In order to properly attach the LVA degradation tag, the prefix of the tag sequence must be complementary to the suffix of Hbif. For this part, the prefix and suffix are reversed in comparison to above: the prefix is RFC25, and the suffix is RFC10, for reasons that are discussed below.
Prepare FimE for protein expression
Team Reports
