Difference between revisions of "Part:BBa K1396000:Experience"
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| + | Expression of BioBrick in conditions outlined above. Row A top represents fluorescence of induced construct. Row A bottom represents optical density over a 16 hour growth period. Row C top represents fluorescence of control, uninduced construct. Row C bottom represent the optical density over a 16 hour growth period. | ||
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Latest revision as of 04:18, 1 November 2014
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K1396000
- Meant for purification and characterization of novel recombinant protein containing an anti-microbial domain and an adhesive peptide domain.
- Contains DNA sequence for MFP-151 recoded with Amber Codons for a L-DOPA orthogonal translation system.
- Antimicrobial peptide, LL-37, has removable N-Terminal purification cap. FLAG tag can be cleaved using enterokinase.
User Reviews
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BBa_K1396000 ahleyva65 |
Team 2014 Yale iGEM ordered this part and attempted to express it within a standard pZE21 plasmid with a pLtetO promoter. Expression may have been successful, but part is extremely toxic and stalls cell growth significantly. In addition, the part is meant for expression in recoded Amberless E. Coli with either a Tyrosine Supressor or L-DOPA Orthogonal Translation System. In the case of L-DOPA the device must expressed in Minimal Media with DTT added one hour before L-DOPA and inducer, although the growth of the E. Coli in Minimal Media was insubstantial for characterization. Growth Kinetic Assay over 16 hours of the construct in pZE21 with atc in LB min with a Tyrosine Supressor indicated a short cycle of growth and death followed by leveling off.
Expression of BioBrick in conditions outlined above. Row A top represents fluorescence of induced construct. Row A bottom represents optical density over a 16 hour growth period. Row C top represents fluorescence of control, uninduced construct. Row C bottom represent the optical density over a 16 hour growth period.
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