Difference between revisions of "Part:BBa K4361116"
 
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<partinfo>BBa_K4361116 short</partinfo>
 
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This part shows the GHB / GBL reporter system designed by the Bielefeld-CeBiTec iGEM 2015 team ([[Part:BBa_K1758376]]) after site-directed mutagenesis with primers [[Part:BBa_K4361112]] and [[Part:BBa_K4361113]], resulting in a deletion in the Blc operator sequence. This mutation is expected to disrupt BlcR binding to the operator, meaning the protein cannot inhibit downstream gene expression. As the remainder of the reporter system remains intact, this part, when inserted in plasmid pSB1C3 (see [[Part:BBa_K4361117]]), may act as a negative control against [[Part:BBa_K4361115]] in experiments where BlcR's binding properties are measured.
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This part shows the GHB / GBL reporter system designed by the Bielefeld-CeBiTec iGEM 2015 team ([[Part:BBa_K1758376]]) but with a deletion in the <i> blc </i> operator sequence. Deletion is inserted with primers [[Part:BBa_K4361112]] and [[Part:BBa_K4361113]].
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It is known from  literature that this mutation disrupts the binding between BlcR and the <i> blc </i> operator sequence [1]. This way BlcR cannot inhibit downstream gene expression.  
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The mutated <i> blc </i> operator sequence is incorperated in a superfolded GFP production plasmid, see[[Part:BBa_K4361115]].
  
 
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===Usage and Biology===
 
===Usage and Biology===
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<h3> Usage and Biology </h3>
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<partinfo>BBa_K4361116 parameters</partinfo>
 
<partinfo>BBa_K4361116 parameters</partinfo>
 
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<h3> References </h3>
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[1] Pan, Y., Fiscus, V., Meng, W., Zheng, Z., Zhang, L.-H., Fuqua, C. and Chen, L. (2011). The Agrobacterium tumefaciens Transcription Factor BlcR Is Regulated via Oligomerization. The Journal of Biological Chemistry, [online] 286(23), pp.20431–20440. doi:10.1074/jbc.M110.196154

Latest revision as of 14:40, 12 October 2022


GHB / GBL biosensor reporter, deletion in Blc operator

This part shows the GHB / GBL reporter system designed by the Bielefeld-CeBiTec iGEM 2015 team (Part:BBa_K1758376) but with a deletion in the blc operator sequence. Deletion is inserted with primers Part:BBa_K4361112 and Part:BBa_K4361113.

It is known from literature that this mutation disrupts the binding between BlcR and the blc operator sequence [1]. This way BlcR cannot inhibit downstream gene expression.

The mutated blc operator sequence is incorperated in a superfolded GFP production plasmid, seePart:BBa_K4361115.

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 121


References

[1] Pan, Y., Fiscus, V., Meng, W., Zheng, Z., Zhang, L.-H., Fuqua, C. and Chen, L. (2011). The Agrobacterium tumefaciens Transcription Factor BlcR Is Regulated via Oligomerization. The Journal of Biological Chemistry, [online] 286(23), pp.20431–20440. doi:10.1074/jbc.M110.196154