Difference between revisions of "Part:BBa K1692004"
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− | [[File:SB2015_PAL_functionality.png|thumbnail|center|500px|<b>PAL | + | [[File:SB2015_PAL_functionality.png|thumbnail|center|500px|<b>PAL Kinetic time course assay</b> Using a Spectramax Pro spectrophotometer we tracked, in real time, the absorbance of trans-cinnamic acid at 268 nm. We ran a total of 24 reactions with varying concentrations of phenylalanine so that we could determine the kinetic parameters of our enzyme. Shown here is a time profile of trans-cinnamic acid production starting with 0.6 mM phenylalanine over a four hour period. Sample points were taken every two minutes. This is where we write]]<br><br> |
<h2>Reference</h2> | <h2>Reference</h2> |
Revision as of 23:46, 17 September 2015
codon optimized PAL with T7 promoter and Flag Tag
Introduction
Phenylalanine ammonia lyase (PAL) catalyzes the conversion of L-phenylalanine to trans-cinnamic acid. Our PAL construct is codon-optimized for expression in E. coli. The original sequence is derived from Anabaena variabilis. We chose the A. variabilis variant of PAL because the literature has characterized it as functioning well, in contrast to University of British Columbia’s 2013 PAL biobrick part (BBa_K1129003) from Streptomyces maritimus, which has much lower activity.
Background
Reference
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1315
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1533
- 1000COMPATIBLE WITH RFC[1000]