Difference between revisions of "Part:BBa K2213004"
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Fusion of the PduD(1-20), Corynebacterium glutamicum class II polyphosphate kinase and mCherry followed by a hexahistidine tag. | Fusion of the PduD(1-20), Corynebacterium glutamicum class II polyphosphate kinase and mCherry followed by a hexahistidine tag. | ||
− | The construct was heterologously expressed in a BL21 (DE3) strain of E. coli under the control of a T7 promoter. The construct was cleaved before or during purification, resulting in the elution of a ~30 kDa, his-tagged protein with a bright pink colour from the Ni-NTA agarose media (<b>Figure 1</b>). | + | The construct was heterologously expressed in a BL21 (DE3) strain of E. coli under the control of a T7 promoter. The construct was cleaved before or during purification, resulting in the elution of a ~30 kDa (expected wieght: 66 kDa), his-tagged protein with a bright pink colour from the Ni-NTA agarose media (<b>Figure 1</b>). The protein fragment was not tested for kinase activity. |
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Revision as of 19:22, 1 November 2017
PduD(1-20)_cgPPK2_mCherry_His6
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Usage and Biology
Fusion of the PduD(1-20), Corynebacterium glutamicum class II polyphosphate kinase and mCherry followed by a hexahistidine tag.
The construct was heterologously expressed in a BL21 (DE3) strain of E. coli under the control of a T7 promoter. The construct was cleaved before or during purification, resulting in the elution of a ~30 kDa (expected wieght: 66 kDa), his-tagged protein with a bright pink colour from the Ni-NTA agarose media (Figure 1). The protein fragment was not tested for kinase activity.
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Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 148
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1030
Illegal SapI.rc site found at 827