Difference between revisions of "Part:BBa K3196011"
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<h1>'''Characterization'''</h1> | <h1>'''Characterization'''</h1> | ||
This is a four section for degrade and transfer lignin part. | This is a four section for degrade and transfer lignin part. | ||
| − | [[File:T--HUST--China--2019-FLO10SLAC.jpg | | + | [[File:T--HUST--China--2019-FLO10SLAC.jpg |350px|thumb|center|Figure1. AOX1-FLO10-SLAC-His tag.]] |
<h1>'''DNA Gel Electrophoretic'''</h1> | <h1>'''DNA Gel Electrophoretic'''</h1> | ||
Revision as of 14:02, 19 October 2019
AOX1-Kozak-FLO10-SLAC-His tag-AOX1 Terminator
FLO10 is a Lectin-like protein with similarity to Flo1 p, thought to be expressed and involved in flocculation. SLAC can catalyze lignin.
Characterization
This is a four section for degrade and transfer lignin part.
DNA Gel Electrophoretic
After we link FLO10 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 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
Illegal BamHI site found at 948 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1387