Part:BBa_K676012:Experience

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Applications of BBa_K676012

According to our result in the 1D chloroquine gel electrophoresis (at 2.5 μg/mL), the plasmid with the pBR322 GBS (on pSB1C3 backbone) did not migrate farther compared to the control plasmid (pSB1C3 with RFP expression cassette - BBa_J04450). At the chloroquine concentration which we used, the more negatively supercoiled plasmid will migrate further while the more relaxed molecules will travel slower through the gel.

Therefore this gel indicate that the pBR322 despite being a gyrase binding site has either very weak affinity for gyrase or is not good at introducing supercoils following the association with gyrase.

Besides achieving a higher level of supercoiling, the quality of the supercoiled plasmids also showed to be better. There seems to be a 34% reduction in the level of maximum supercoiling in the plasmid with pBR322 when compared to the control. Also there is a reduction in the consistency of supercoiling with the pBR322 GBS as displayed by the wider range of the band for the sample.

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