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:Kozak_sequence.png ‎|400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]]
+
[[ 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.

Figure1. T--HUST--China--2019-SUC2VP

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.

Figure1. This figure shows the most possible kozak consensus sequence.

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.

Figure1. This figure shows the most possible kozak consensus sequence.

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.

Figure1. This figure shows the most possible kozak consensus sequence.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 937
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]