Difference between revisions of "Part:BBa K3196025"
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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: | + | [[ File:T--HUST--China--2019-SUC2VP.jpg|400px|thumb|center|Figure1. T--HUST--China--2019-SUC2VP]] |
<h1>'''DNA Gel Electrophoretic'''</h1> | <h1>'''DNA Gel Electrophoretic'''</h1> | ||
Revision as of 15:46, 19 October 2019
AOX1-Kozak-SUC2-VP-His tag-AOX1 Terminator
The SUC2 gene of S. cerevisiae, encoding invertase was designed to codon preference of P. pastoris excluding the sequence of the N-terminal pre-pro-sequence signal peptide and His-tag in the C terminus. VP can catalyze pectin.
Characterization
This is a four section for degrade and transfer lignin part.
DNA Gel Electrophoretic
After we link SUC2 and VP successfully, we run the PCR with an intention to confirm the expression of VP. 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 SUC2 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 manganese and hydrogen peroxide to detect the enzyme activity. As the figure shows, the solution turns xxx, 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]
- 1000COMPATIBLE WITH RFC[1000]