Difference between revisions of "Part:BBa K2213004"

Revision as of 00:50, 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 a ~30 kDa, his-tagged protein with a bright pink colour (figure 1). Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 148
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1030
Illegal SapI.rc site found at 827

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-This part is a fusion of the PduD(1-20), cgPPK and mCherry followed by a hexahistidine tag. We intended the PduD(1-20) sequence to localise the cgPPK into a recombinant microcompartment and the mCherry would have allowed us to confirm its co-localisation in the microcompartment.
+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 a ~30 kDa, his-tagged protein with a bright pink colour (figure 1).
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 <span class='h3bb'>Sequence and Features</span>