Part:BBa_K1149008
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 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 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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
Illegal NheI site found at 1009 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 356
Illegal BamHI site found at 65 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 210
Illegal AgeI site found at 1531 - 1000COMPATIBLE WITH RFC[1000]
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