Part:BBa_K3711066

Pynr071c-α factor-DsbC-AOX1 Terminator

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1438
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1593
  
• 1000
  COMPATIBLE WITH RFC[1000]

Description

This is a composite part for extracellular expression of protein disulfide isomerase(PDI). Pynr071c is the promoter of Aldose 1-isomerase superfamily protein YNR071C. In the presence of xylose, the signal peptide, α-factor , is used to express DsbC outside of the cell.

Usage and Biology

DsbC is one of five essential protein needed for forming disulfide bonds in E.coli, functioning as disulfide bond isomerase during folding process of oxidized protein in periplasmic space. DsbC is a homodimer of size of 2x23kDa, with enzymatic activity of protein disulfide bonds isomerase and molecular chaperone.
The crystal structure of DsbC shows that disulfide bonds are formed in all Cys-X-X-Cys reaction center. Through the hinge, which allows the reaction center moving relatively, a single thioredoxin-like domain is linked to dimer domain in the amino-terminal. The widely-existing uncharged seam between reaction center may participate in peptide combination and enzymatic reaction of DsbC folding enzymes.
Linking DsbC with peptides rich in cysteine, we build a transporting and auxiliary binding complex with disulfide bond isomerase and short peptides.

Molecular cloning

Fig1. Colony PCR results of AOX1-α factor-curA-AOX1 Terminator, AOX1-α factor-pepACS-AOX1 Terminator and AOX1-α factor-DsbC-AOX1 Terminator transformed E.coli