Difference between revisions of "Part:BBa K2997009"
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===Expression in <i>E.coli</i>=== | ===Expression in <i>E.coli</i>=== | ||
− | + | We constructed Tyrosine Ammonia Lyase with a native RBS , and transformed the plasmid into E. coli Nissle 1917 and confirmed it by double digestion. The results are as follows: | |
Revision as of 17:26, 19 October 2019
J23100-NRBS-sam8 (J23100-I742106)
Background
Tyrosine ammonia-lyase (TAL) is an enzyme that converts tyrosine into p-Coumaric acid. We engineered E. coli Nissle to express TAL to turn the excess tyrosine inside the gut into p-Coumaric acid, and compare the different constructs for improvement, BBa_K2997009 and BBa_K2997010.
Expression in E.coli
We constructed Tyrosine Ammonia Lyase with a native RBS , and transformed the plasmid into E. coli Nissle 1917 and confirmed it by double digestion. The results are as follows:
Fig. 6. Confirmation of BBa_K2997009 by double digestion, arrow indicates TAL w/ NRBS (~1600bp).M: Marker; Lane 1: pSB1C3-BBa_K2997009; Lane 2: BBa_K2997009
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 457
Illegal BamHI site found at 36 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 232