Part:BBa_K652000
T7-RBS-eGFP-TEV-RiAFP-His-Term
This part contains a T7 strong promoter, RBS, our eGFP-fusion protein containing the Rhagium inquistor antifreeze protein (RiAFP) with a His tag at the C-terminus, followed by a terminator.
More information about this and other parts can be found on our wiki under the project section: http://2011.igem.org/Team:Yale
Cloning results were confirmed by colony PCR, restriction digest, and full sequencing.
Expression levels of up to 150mg/L of this protein are detected by UV-vis.
Figure 1: Expression of RiAFP-GFP for purification. File:Jen RiGFP Purification (08-04-2011).jpg
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Figure 2-3: Cleavage at TEV site for isolation of RiAFP
Figure 4: RiAFP-GFP increases survivability of E. coli
Figure 5: RiAFP-GFP inhibits ice recrystallization
Figure 6: RiAFP-GFP enhances
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 894
Illegal AgeI site found at 990
Illegal AgeI site found at 1104 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 74
Illegal SapI.rc site found at 87
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