Difference between revisions of "Part:BBa K3196012"
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<partinfo>BBa_K3196012 short</partinfo> | <partinfo>BBa_K3196012 short</partinfo> | ||
− | protein | + | FLO10 αpro is a combined signal peptide, which enhance the enzyme activity 3 times. SLAC can catalyze lignin. |
+ | <h1>'''Characterization'''</h1> | ||
+ | This is a four section for degrade and transfer lignin part. | ||
+ | [[File:Kozak_sequence.png |400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]] | ||
+ | |||
+ | <h1>'''DNA Gel Electrophoretic'''</h1> | ||
+ | After we link FLO10 αpro and SLAC successfully, we run the PCR with an intention to confirm the expression of SLAC. As the figure shows, we get the genetic stripe about xxx bp which means the PCR is successful. | ||
+ | [[File:Kozak_sequence.png |400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]] | ||
+ | |||
+ | <h1>'''SDS-PAGE'''</h1> | ||
+ | We run the SDS-PAGE to check whether FLO10 αpro help the enzyme transfer to the extracellular. As the figure shows, we get the protein type about xxx bp which means the SDS-PAGE is successful. | ||
+ | [[File:Kozak_sequence.png |400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]] | ||
+ | |||
+ | <h1>'''Enzyme Activity'''</h1> | ||
+ | We use ABTS to detect the enzyme activity. As the figure shows, the solution turns green, which confirm the enzyme activity. | ||
+ | [[File:Kozak_sequence.png |400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]] | ||
− | |||
− | |||
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Revision as of 11:27, 19 October 2019
AOX1-Kozak-FLO10 pro-SLAC-His tag-AOX1 Terminator
FLO10 αpro is a combined signal peptide, which enhance the enzyme activity 3 times. SLAC can catalyze lignin.
Characterization
This is a four section for degrade and transfer lignin part.
DNA Gel Electrophoretic
After we link FLO10 αpro and SLAC successfully, we run the PCR with an intention to confirm the expression of SLAC. As the figure shows, we get the genetic stripe about xxx bp which means the PCR is successful.
SDS-PAGE
We run the SDS-PAGE to check whether FLO10 αpro help the enzyme transfer to the extracellular. As the figure shows, we get the protein type about xxx bp which means the SDS-PAGE is successful.
Enzyme Activity
We use ABTS to detect the enzyme activity. As the figure shows, the solution turns green, which confirm the enzyme activity.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 937
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1586