Generator
AntiFreeze

Part:BBa_K193209:Experience

Designed by: Nao Nakatani   Group: iGEM09_Tokyo_Tech   (2009-10-14)
Revision as of 17:41, 23 September 2011 by AaronHakim1991 (Talk | contribs) (→‎Applications of BBa_K193209)

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Please enter how you used this part and how it worked out.

Applications of BBa_K193209

Team Yale has submitted a sequence of this biobrick that is BBF RFC10 compatible (BBa_K652002 and BBa_K652003). See: https://parts.igem.org/wiki/index.php?title=Part:BBa_K652002.

Our biobrick contains a T7 strong promoter, RBC, eGFP, followed by the Tenebrio molitor antifreeze protein (TmAFP) with a His tag at the N-terminus, followed by a terminator. Unlike the Tokyo Tech TmAFP biobrick, ours does not contain an internal EcoRI restriction site. Additionally, BBa_K652002 has a TEV protease linker between the eGFP and the TmAFP which allows for isolation of the TmAFP part.

We kindly obtained the sequence of the TmAFP from the Fass Lab, reported on in the following paper:

Bar, M., Bar-Ziv, R., Scherf, T. & Fass, D. Efficient production of a folded and functional, highly disulfide-bonded [beta]-helix antifreeze protein in bacteria. Protein Expression and Purification 48, 243-252 (2006).

The sequence of the Tokyo Tech part BBa_K193209 seems to have part of the TmAFP protein missing.

User Reviews

UNIQ10e094870bb65d8f-partinfo-00000000-QINU UNIQ10e094870bb65d8f-partinfo-00000001-QINU