Difference between revisions of "Part:BBa K3196013"
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<partinfo>BBa_K3196013 short</partinfo> | <partinfo>BBa_K3196013 short</partinfo> | ||
− | + | PHO1 is one of three repressible acid phosphatases, a glycoprotein that is transported to the cell surface. 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 PHO1 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 PHO1 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 11:30, 19 October 2019
AOX1-Kozak-PHO1-SLAC-His tag-AOX1 Terminator
PHO1 is one of three repressible acid phosphatases, a glycoprotein that is transported to the cell surface. SLAC can catalyze lignin.
Characterization
This is a four section for degrade and transfer lignin part.
DNA Gel Electrophoretic
After we link PHO1 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 PHO1 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 1367