DNA

Part:BBa_K676011

Designed by: Meng Li   Group: iGEM11_UCL_London   (2011-09-19)

Gyrase Binding Site from pSC101

The gyrase binding site cloned from pSC101 plasmid.

Usage and Biology

The pSC101 GBS has highly efficient binding to GBS and yet lacks the stimulation of supercoiling (Oram, 2003) and this raises the possibility that the biological effects of gyrase at par are purely structural, for example, local changes in DNA topology due to the wrapping of DNA by gyrase may be already enough to induce the downstream effects of gyrase on pSC101 replication and partition.

Subsequent studies concluded that the key function of par was to cause a local increase in superhelical density in the origin region of pSC101 (Conley and Cohen, 1995), and this in turn was critical in determining the interactions of other replication and partition proteins within the origin itself (Miller and Cohen, 1999). However, further experiments will be necessary to address specific questions relating to the role of gyrase function at par at a more detailed or molecular level.

From the studies, it was also found that the pSC101 GBS is cleaved by gyrase much more efficiently than the pBR322 GBS site, but on the other hand plasmid with pBR322 GBS was supercoiled much better than the one with pSC101 GBS. But still the best supercoiling efficiency was achieved by the plasmid with Mu phage GBS.

Characterising Part BBa K676011.jpg

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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Categories
Parameters
n/aGyrase Binding Site from pSC101