Difference between revisions of "Part:BBa K1149008"

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https://static.igem.org/mediawiki/2013/thumb/a/ab/PK_pelB.png/450px-PK_pelB.png
 
https://static.igem.org/mediawiki/2013/thumb/a/ab/PK_pelB.png/450px-PK_pelB.png
  
'''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.
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'''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.
  
 
https://static.igem.org/mediawiki/2013/thumb/5/54/PK_pelB_fluor.png/450px-PK_pelB_fluor.png
 
https://static.igem.org/mediawiki/2013/thumb/5/54/PK_pelB_fluor.png/450px-PK_pelB_fluor.png
  
'''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.
+
'''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.
  
 
<b>Conclusion: There is no growth inhibition caused by induction, however induction is significantly induced by the Xylose promoter. </b>
 
<b>Conclusion: There is no growth inhibition caused by induction, however induction is significantly induced by the Xylose promoter. </b>

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.

450px-PK_pelB.png

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.

450px-PK_pelB_fluor.png

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


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