Difference between revisions of "Part:BBa K1804002"
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===Usage and Biology===
 
===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 (p''nirB''), which drives the expression of NADH-dependent nitrite reductase (Jayaraman et al., 1987). p''nirB'' 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). p''nirB'' 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, p''nirB'' can be regulated by FNR protein in this system.
 
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 (p''nirB''), which drives the expression of NADH-dependent nitrite reductase (Jayaraman et al., 1987). p''nirB'' 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). p''nirB'' 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, p''nirB'' can be regulated by FNR protein in this system.
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===Part Submitted===
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We have sequenced the part we submitted, and found that it contains the ''nirB'' promoter and the B0032 ribosome binding site, but not the full invasin sequence. The BioBricks Prefix and Suffix are present.
  
 
===References===
 
===References===

Revision as of 13:09, 21 September 2015

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.

Part Submitted

We have sequenced the part we submitted, and found that it contains the nirB promoter and the B0032 ribosome binding site, but not the full invasin sequence. The BioBricks Prefix and Suffix are present.

References

Griffiths, L., & Cole, J. A. (1987). Lack of redox control of the anaerobically-induced nirB+ gene of Escherichia coli K-12. Archives of microbiology, 147(4), 364-369.

Jayaraman, P. S., Peakman, T. C., Busby, S. J. W., Quincey, R. V., & Cole, J. A. (1987). Location and sequence of the promoter of the gene for the NADH-dependent nitrite reductase of Escherichia coli and its regulation by oxygen, the Fnr protein and nitrite. Journal of molecular biology, 196(4), 781-788.

Spiro, S., & Guest, J. R. (1987). Regulation and over-expression of the fnr gene of Escherichia coli. Journal of general microbiology, 133(12), 3279-3288.


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]