Part:BBa_K2812002

pBAD-ara promoter - Arabinose inducible regulatory promoter

The pBAD promoter is part of the arabinose operon. A gene under the control of the pBAD promoter can be induced by high arabinose concentrations. We as iGem Eindhoven 2018 used this promoter to control the release of the antimicrobials lysostaphin and pyocin.

Protein Expression

Figure 1: SDS-PAGE gel of pBAD promotor-Lysostaphin-thrombin-HlyA-His-tag construct protein expression experiment.

Protein expression experiments were performed to characterize the functionality of the pBAD promotor. To allow expression under the control of the construct by the addition of arabinose, it was assembled to the BBa_K2812004 biobrick. The biobrick was successfully transformed into BL21 (DE3), after which a culture was set up at 37 °C. A sample was taken before induction (0). A culture was induced at an OD600 of 0.5-0.8 by adding 2 mM arabinose to induce expression of the recombinant protein, also at 37 °C. Samples were taken 4 hours after induction of both an induced (4+) and an uninduced culture (4-). SDS samples were prepared and loaded onto a polyacrylamide gel to yield the SDS-PAGE results that can be seen in figure 1. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Unknown
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]