Difference between revisions of "Part:BBa K4307008"
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| − | + | Originated from <i>Lactococcus lactis subsp. Lactis</i>, NisR is a member of the two-component regulatory system NisK/NisR involved in the regulation of the biosynthesis of lantibiotic nisin. NisR functions as a regulatory protein which is phosphorylated by NisK in response to environmental nisin signals. We managed to express NisR in <i>E.coli</i> strain.<br> | |
| + | Functions together with NisK(BBa_K4307007), PnisA(BBa_K4307009), J23100 and EGFP, it forms composite part J23100-nisK-nisR+PnisA-EGFP(BBa_K4307013) to express fluorescence signal induced by nisin. | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
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| − | < | + | <h2>Characterization</h2> |
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| + | <p>The following figure demonstrates our successful construction.</p > | ||
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| + | <br> | ||
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| + | https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307008-gel.png<br> | ||
| + | <b>Figure 1: </b> <b>The construction results of NisR.</b> | ||
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| + | <h3>SDS-PAGE assay was done to characterize the biobrick.</h3> | ||
| + | <p>To determine the expression of NisR, we used T7 promoter and lac operator to control the overexpression of NisR by IPTG. NisR in bacterial lysate was detected by SDS-PAGE(Figure 2), indicating that NisR can be successfully expressed in <i>E.coli</i>. </p > | ||
| + | <p> | ||
| + | <br> | ||
| + | https://static.igem.wiki/teams/4307/wiki/wiki-parts-files/mainpage/bba-k4307008.png<br> | ||
| + | <b>Figure 2: </b> <b> SDS-PAGE result of NisR expression.</b> | ||
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| + | </p > | ||
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| + | <h2>Conclusion</h2> | ||
| + | <p>According to the SDS-PAGE result, it can be found that NisR can be successfully expressed in <i>E.coli</i>, which indicates that it can act as the downstream regulator of NisK to pass down nisin signal functionally in <i>E.coli</i> as in <i>Lactococcus lactis</i>. This ensures the effectiveness of nisin TCS in <i>E.coli</i>.</p > | ||
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| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K4307008 SequenceAndFeatures</partinfo> | ||
Revision as of 15:11, 12 October 2022
NisR
Originated from Lactococcus lactis subsp. Lactis, NisR is a member of the two-component regulatory system NisK/NisR involved in the regulation of the biosynthesis of lantibiotic nisin. NisR functions as a regulatory protein which is phosphorylated by NisK in response to environmental nisin signals. We managed to express NisR in E.coli strain.
Functions together with NisK(BBa_K4307007), PnisA(BBa_K4307009), J23100 and EGFP, it forms composite part J23100-nisK-nisR+PnisA-EGFP(BBa_K4307013) to express fluorescence signal induced by nisin.