Part:BBa_K4788008

FadR-FadBA-FadO-mRFP

This is an oleic acid inducer connected to the mRFP protein to characterize the effect of the oleic acid inducer.

Usage and Biology

The entire part is formed by adding the mRFP protein gene to the oleic acid inducer, which is used to characterize the oleic acid inducer. When the oleic acid inducer is activated, the mRFP protein will be expressed, and we can use fluorescence intensity to measure the expression level of the oleic acid inducer.

Characterization

To verify the effect of oleic acid inducer, we added mRFP gene after the oleic acid inducer gene to construct a reporter gene. We cultured engineering bacteria under two different environmental conditions, anaerobic and aerobic, and under different oleic acid induction conditions, and detected the expression effect of oleic acid inducers through normalized fluorescence signal intensity. As shown in Figure 1, the oleic acid inducer has a good response to gradient oleic acid, especially in the absence of oxygen, showing significant differences from the previous gradient at a 10% oleic acid gradient.

< img style="max-width:700px;" src="https://static.igem.wiki/teams/4788/wiki/proof/fad-separated.jpg" alt="control">

Figure 1: Gradient oleic acid induced expression intensity of oleic acid starter under both anaerobic and aerobic conditions

Under the comparison of anaerobic and aerobic conditions, as shown in Figure 2, there was a slight difference in the expression level of the oleic acid inducer gene, but it did not show significant difference.

< img style="max-width:700px;" src="https://static.igem.wiki/teams/4788/wiki/proof/fad-all.png" alt="control">

Figure 2: Comparison of the expression intensity of gradient oleic acid induced oleic acid starter under anaerobic and aerobic conditions

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 764
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1343
  Illegal AgeI site found at 1455
  
• 1000
  COMPATIBLE WITH RFC[1000]