Part:BBa_K1149007
Proteinase K
Encodes a peptidase from proteinase K family, breaks down polylactic acid (no PelB secretion tag). The BBa_K1149008 construct is the same enzyme but with the pelB secretion tag.
Characterisation
Our Proteinase K (no 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 Arabinose.
Proteinase K growth assay. MG1655 E. coli display strong growth with the induction of BBa_K1149007, though it is not significant as a two-tailed t-test gave p = 0.1471. Growth was at 37°C with shaking. Error bars are SEM, n=4. Figure made by Imperial College London 2013 iGEM.
Proteinase K induction assay. When induced, there is a very clear increase in fluorescence for BBa_K1149007, however t-test analysis gives p = 0.0595 > 0.05 for this, thus the fluorescence does not significantly differ from uninduced. 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, but fluorescence from induction is elevated in Proteinase K (intracellular).
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 943 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 290
Illegal BamHI site found at 65 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1465
- 1000COMPATIBLE WITH RFC[1000]
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