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.  
  
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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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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>).
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<img src= "https://static.igem.org/mediawiki/2017/8/8f/PduD%281-20%29_cgPPK_mCherry_%281%29.png" width="800" height="600">
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<p><font color="#111"><center></h3><b>Figure 1.</b> SDS-PAGE analysis of fractions from the Ni-NTA agarose purification of the
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 17:13, 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, his-tagged protein with a bright pink colour from the Ni-NTA agarose media (Figure 1).
</br> <img src= "PduD%281-20%29_cgPPK_mCherry_%281%29.png" width="800" height="600">

</h3>Figure 1. SDS-PAGE analysis of fractions from the Ni-NTA agarose purification of the

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