Part:BBa_K3196020

AOX1-Kozak-SUC2-pelA-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. pelA can catalyze pectin.

Characterization

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

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

DNA Gel Electrophoretic

After we link SUC2 and pelA successfully, we run the PCR with an intention to confirm the expression of pelA. 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 DNS 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 BglII site found at 1761
  Illegal BamHI site found at 937
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]