Difference between revisions of "Part:BBa K3196015"
 
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<partinfo>BBa_K3196015 short</partinfo>
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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. SLAC can catalyze lignin.
 
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. SLAC can catalyze lignin.
  
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[[File:T--HUST--China--2019-SUC2SLAC.jpg ‎|400px|thumb|center|Figure1. T--HUST--China--2019-SUC2SLAC]]
 
[[File:T--HUST--China--2019-SUC2SLAC.jpg ‎|400px|thumb|center|Figure1. T--HUST--China--2019-SUC2SLAC]]
  
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<h1>'''DNA Gel Electrophoretic'''</h1>
 
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After we link SUC2 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.
 
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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 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.
 
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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 ABTS to detect the enzyme activity. As the figure shows, the solution turns green, 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.]]
 
  
  

Latest revision as of 07:39, 20 October 2019


AOX1-Kozak-SUC2-SLAC-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. SLAC can catalyze lignin.

Characterization

This is a four section for degrade and transfer lignin part.

Figure1. T--HUST--China--2019-SUC2SLAC



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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1361