Part:BBa_K676001

Gyrase Binding Site from Mu Bacteriophage

The strong gyrase site cloned from Mu bacteriophage genome.

Usage and Biology

The Mu phage strong gyrase site (SGS) serves as a high affinity binding site for the DNA gyrase enzyme. This site is known to be cleaved preferentially by the Gyrase A subunits and therefore results in an increase in the linking number difference of the DNA molecule.

Apparently, some bacteriophages are known to exploit the GBS to allow them to replicate successfully in their host. For example, Mu bacteriophage has a strong gyrase site or GBS in the central region of their 37.2 kbp genome. From work carried out by Pato et al. in 1990, it was found out that the wild type phage can carry out replication effectively compared a mutant which lacks the GBS after the heat induction of the lysogens.

By introducing the Mu GBS into the target plasmids, we managed to achiev a higher level of supercoiling as well as better uniformity in the final supercoiled products. Highly supercoiled plasmids are able to resist shear stress better and this property plays an important role in large scale plasmid DNA manufacturing as well as for the delivery therapy of therapeutic genes.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 191
  
• 1000
  COMPATIBLE WITH RFC[1000]