Difference between revisions of "Part:BBa K3408000"

Revision as of 16:58, 27 October 2020

promoter nar(fnr regulated)

P_nar is an oxygen-free inducible promoter which can response to the ResD/E two-component regulatory system and the global regulator Nitrate Reductase Regulator (FNR) of Bacillus subtilis.

In our project, we use it to express Phy(ycD) efficiently in the absence of oxygen, and meanwhile express CI repressor protein to inhibit toehold switch-mzaF downstream.

According to the paper, Pnar with three levels of strength were designed and applied in E.coli successfully. The expression level of GFP in the control of high-strength promoter was 19.7 higher than the wild-type promoter.

Fig.1. The spacer sequence of 15 bp between the -35 and -10 elements of the upstream region of the wild-type nar promoter was randomized(Hwang Hee Jin, et al., BIOTECHNOL BIOFUELS. 2018)

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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-We add the promoter P<sub>nar</sub>, <b>in response to the RES-FNR oxygen regulator system</b> of <i>Bacillus Subtilis</i>. It is activated by ResD which is phosphorylated by ResE when there is no oxygen. In our project, we use it to express <b>Phy(ycD)</b> efficiently in the absence of oxygen, and meanwhile express <b>CI repressor protein</b> to inhibit toehold switch-mzaF downstream.
+P<sub>nar</sub> is an oxygen-free inducible promoter which can response to the ResD/E two-component regulatory system and the global regulator Nitrate Reductase Regulator (FNR) of <i>Bacillus subtilis</i>.
+In our project, we use it to express <b>Phy(ycD)</b> efficiently in the absence of oxygen, and meanwhile express <b>CI repressor protein</b> to inhibit toehold switch-mzaF downstream.
 According to the paper, Pnar with three levels of strength were designed and applied in <i>E.coli</i> successfully. The expression level of GFP in the control of high-strength promoter was 19.7 higher than the wild-type promoter.