Difference between revisions of "Part:BBa K1692004"
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− | [[File:SB2015_PAL_initial test.png|thumbnail|center|900px|< | + | [[File:SB2015_PAL_initial test.png|thumbnail|center|900px|<b>In an initial assay</b>, we took absorbance spectra of the following reactions using a Nanodrop 2000 machine. <b>[Left]</b> Phenylalanine and PAL separate. A negative control to show the absorbance of pure phenylalanine and pure PAL in tris buffer (ph 8.0). |
− | [Center] Phenylalanine and PAL together. We observed a very large absorbance peak at 268 nm, suggesting that trans-cinnamic acid was produced. | + | <b>[Center]</b> Phenylalanine and PAL together. We observed a very large absorbance peak at 268 nm, suggesting that trans-cinnamic acid was produced. |
− | [Right] Pure trans-Cinnamic Acid. A positive control to confirm the absorbance peak of pure trans-cinnamic acid. | + | <b>[Right]</b> Pure trans-Cinnamic Acid. A positive control to confirm the absorbance peak of pure trans-cinnamic acid. |
]]<br><br> | ]]<br><br> | ||
Revision as of 22:58, 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]