Difference between revisions of "Part:BBa K1804002"
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In this construct, the invasin gene from Yersina is placed under the expression of the anaerobic-sensitive promoter pnirB from Escherichia coli. | In this construct, the invasin gene from Yersina is placed under the expression of the anaerobic-sensitive promoter pnirB from Escherichia coli. | ||
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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, pnirB can be regulated by FNR protein in this system. | ||
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Revision as of 08:28, 20 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.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal NotI site found at 7 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
- 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal XbaI site found at 16 - 1000COMPATIBLE WITH RFC[1000]