Difference between revisions of "Part:BBa K1149002"
Margarita K (Talk | contribs) |
Margarita K (Talk | contribs) |
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<partinfo>BBa_K1149002 short</partinfo> | <partinfo>BBa_K1149002 short</partinfo> | ||
| − | This part expresses a plastic degradation enzyme and | + | This part expresses a plastic degradation enzyme and [https://parts.igem.org/Part:BBa_K515005 sfGFP] under the control of arabinose inducible promoter. |
The enzyme is the estCS2 from [https://parts.igem.org/Part:BBa_K892012 BBa_K892012] previous biobrick by Washington 2012 team, that encodes carboxylesterase that breaks down ester bonds in polyurethane. We have added a [https://parts.igem.org/Part:BBa_K1149023 pelB] seretion tag to the enzyme so that it can be secreted extacellularly. We also added a 6xHistidine tag for Western blotting and protein purification purposes. | The enzyme is the estCS2 from [https://parts.igem.org/Part:BBa_K892012 BBa_K892012] previous biobrick by Washington 2012 team, that encodes carboxylesterase that breaks down ester bonds in polyurethane. We have added a [https://parts.igem.org/Part:BBa_K1149023 pelB] seretion tag to the enzyme so that it can be secreted extacellularly. We also added a 6xHistidine tag for Western blotting and protein purification purposes. | ||
Revision as of 17:27, 15 September 2013
pelB-EstCS2-his expression (GFP reporter)
This part expresses a plastic degradation enzyme and sfGFP under the control of arabinose inducible promoter.
The enzyme is the estCS2 from BBa_K892012 previous biobrick by Washington 2012 team, that encodes carboxylesterase that breaks down ester bonds in polyurethane. We have added a pelB seretion tag to the enzyme so that it can be secreted extacellularly. We also added a 6xHistidine tag for Western blotting and protein purification purposes.
Theoretical pathway of PUR degradation by esterase:
references:
[http://www.selu.edu/acad_research/depts/biol/faculty/publications/pdf/2012/howard2012.pdf Howard, Gary 2012 pg:371 -3, Polyurethane Biodegradation, Environmental Science]
[http://www.microbialcellfactories.com/content/10/1/41 A novel family VII esterase with industrial potential from compost metagenomic library. Microbial Cell Factories 2011]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
Illegal BamHI site found at 692 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 210
Illegal NgoMIV site found at 958
Illegal NgoMIV site found at 1192
Illegal AgeI site found at 2131 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1006