Difference between revisions of "Part:BBa K3196021"
 
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<partinfo>BBa_K3196021 short</partinfo>
 
<partinfo>BBa_K3196021 short</partinfo>
  
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FLO10 is a Lectin-like protein with similarity to Flo1 p, thought to be expressed and involved in flocculation. VP can catalyze pectin.
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FLO10 is a Lectin-like protein with similarity to Flo1 p, thought to be expressed and involved in flocculation. VP can catalyze lignin.
  
 
<h1>'''Characterization'''</h1>
 
<h1>'''Characterization'''</h1>
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This is a four section for degrade and transfer lignin part.
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This is a four section for degrade and transfer lignin part.FLO10 is a kind of signal peptide which was initially found before the protein involved in flocculation. It is supposed that the signal peptides from pichia pastoris can be better identified by its signal recognition system, thus benefiting the secretion of exogenous protein. So we add the coding sequence of FLO10 before VP to figure out whether it could increase the output of VP.
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[[File:Kozak_sequence.png ‎|400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]]
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[[File:T--HUST--China--2019-FLO10VP.jpg ‎|400px|thumb|center|Figure1. T--HUST--China--2019-FLO10VP]]
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<h1>'''DNA Gel Electrophoretic'''</h1>
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After we link FLO10 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.
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[[File:Kozak_sequence.png ‎|400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]]
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<h1>'''SDS-PAGE'''</h1>
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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.
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[[File:Kozak_sequence.png ‎|400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]]
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<h1>'''Enzyme Activity'''</h1>
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We use manganese and hydrogen peroxide to detect the enzyme activity. As the figure shows, the solution turns xxx, which confirm the enzyme activity.
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[[File:Kozak_sequence.png ‎|400px|thumb|center|Figure1. This figure shows the most possible kozak consensus sequence.]]
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Latest revision as of 10:36, 20 October 2019


AOX1-Kozak-FLO10-VP-His tag-AOX1 Terminator

FLO10 is a Lectin-like protein with similarity to Flo1 p, thought to be expressed and involved in flocculation. VP can catalyze lignin.

Characterization

This is a four section for degrade and transfer lignin part.FLO10 is a kind of signal peptide which was initially found before the protein involved in flocculation. It is supposed that the signal peptides from pichia pastoris can be better identified by its signal recognition system, thus benefiting the secretion of exogenous protein. So we add the coding sequence of FLO10 before VP to figure out whether it could increase the output of VP.

Figure1. T--HUST--China--2019-FLO10VP

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
    Illegal BamHI site found at 948
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]