Part:BBa_K1804002

pNirB-Invasin

In this construct, the invasin gene from Yersina is placed under the expression of the anaerobic-sensitive promoter pnirB from Escherichia coli.

Usage and Biology

In the facultative anaerobe Escherichia coli K-12, the transcription factor Fumarate and Nitrate reductase Regulatory (FNR) protein is encoded by the fnr gene (Spiro & Guest, 1987). FNR activates various fnr-dependent promoters in the anaerobic pathway of E. coli (Spiro & Guest, 1987), such as the nirB promoter (pnirB), which drives the expression of NADH-dependent nitrite reductase (Jayaraman et al., 1987). pnirB has partial activity when induced under anaerobic conditions and optimal activity in the presence of nitrite in addition to anaerobic growth conditions (Griffiths & Cole, 1987; Jayaraman et al., 1987). pnirB is a switch that can induce the expression of target genes under anaerobic conditions. As the non-pathogenic E. coli K-12 strain BL21 [http://www.ncbi.nlm.nih.gov/nuccore/CP010816.1] (Life Technologies) that we used expresses the fnr gene endogenously, pnirB can be regulated by FNR protein in this system.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 1
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1
  Illegal NotI site found at 7
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 1
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 1
  Illegal XbaI site found at 16
  
• 1000
  COMPATIBLE WITH RFC[1000]