Difference between revisions of "Part:BBa K3576001"
LucyShi 2018 (Talk | contribs) |
LucyShi 2018 (Talk | contribs) |
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===Results=== | ===Results=== | ||
==1. Protein expression test== | ==1. Protein expression test== | ||
| − | SDS-PAGE electrophoresis was used to check the expression of PETase protein. As shown in Figure 1, compared to the blank control, the lane contained PETase ( | + | SDS-PAGE electrophoresis was used to check the expression of PETase protein. As shown in Figure 1, compared to the blank control, the lane contained PETase (40 KDa) protein indicated that this protein has been successfully expressed. |
[[File: PETase-Fig1.png|200px|thumb|center|Figure 1 Protein SDS-PAGE electrophoresis results of PETase.]] | [[File: PETase-Fig1.png|200px|thumb|center|Figure 1 Protein SDS-PAGE electrophoresis results of PETase.]] | ||
Revision as of 21:17, 24 October 2020
PETase expression system
1
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 97
Illegal NgoMIV site found at 123 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 267
Results
1. Protein expression test
SDS-PAGE electrophoresis was used to check the expression of PETase protein. As shown in Figure 1, compared to the blank control, the lane contained PETase (40 KDa) protein indicated that this protein has been successfully expressed.
2. Enzyme Activity Test of PETase
The enzyme activity of PETase was performed by p-NP assay which is a common way to quantify hydrolytic activity. We selected p-Nitrophenylbutyrate (pNPB) as the substrate, which can be hydrolyzed to p-nitrophenol (pNP) (Figure 2-A). The concentration of pNP can be measured by the characteristic absorption at 405 nm. As shown in Figure 2-B, with the extension of reaction time, the OD405 value of p-NP gradually increased, which indicates that the degradation activity of the PETase.
