Difference between revisions of "Part:BBa K1149008"

Revision as of 23:42, 2 October 2013

Proteinase K (extracellular)

Encodes a peptidase from proteinase K family, breaks down polylactic acid. See here for construct with secretion tag.

Characterisation

Our Proteinase K (with PelB secretion tag) construct contains sfGFP within an operon and therefore fluorescence can be utilised to determine if expression is being induced by addition of Xylose.

Proteinase K with pelB secretion tag growth assay. As with Proteinase K without pelB, there is increased growth compared to uninduced for BBa_K1149008. However this is not significant as a two-tailed t-test gave p = 0.4442. Growth was at 37°C in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.

Proteinase K with pelB secretion tag induction assay. Induction of BBa_K1149008 gives a very strong induction response, this is significant as a two-tailed t-test gave a value of p = 0.0104 < 0.05. Thus the induction does have a significant effect on fluorescence for Proteinase K with pelB. Growth was at 37°C in LB with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.

Conclusion: There is no growth inhibition caused by induction, however induction is significantly induced by the Xylose promoter.

References:

Larsen AN, Moe E, Helland R, Gjellesvik DR, Willassen NP.Characterization of a recombinantly expressed proteinase K-like enzyme from a psychrotrophic Serratia sp. FEBS J. 2006 Jan;273(1):47-60.

Sequence and Features