Difference between revisions of "Part:BBa K2213004"
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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 (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.
 
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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https://static.igem.org/mediawiki/2017/7/76/PduD%281-20%29_cgPPK_mCherry_%284%29.png
 
https://static.igem.org/mediawiki/2017/7/76/PduD%281-20%29_cgPPK_mCherry_%284%29.png
 
<p><font color="#111"><center></h3><b>Figure 1.</b> SDS-PAGE analysis of fractions from the Ni-NTA agarose purification of the PduD(1-20)_cgPPK_mCherry construct. The black arrow indicates where the expected band should be (66 kDa) in the elution fraction lane. Ladder: Precision Plus Protein™ (Biorad)
 
<p><font color="#111"><center></h3><b>Figure 1.</b> SDS-PAGE analysis of fractions from the Ni-NTA agarose purification of the PduD(1-20)_cgPPK_mCherry construct. The black arrow indicates where the expected band should be (66 kDa) in the elution fraction lane. Ladder: Precision Plus Protein™ (Biorad)

Revision as of 19:34, 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.

PduD%281-20%29_cgPPK_mCherry_%284%29.png

</h3>Figure 1. SDS-PAGE analysis of fractions from the Ni-NTA agarose purification of the PduD(1-20)_cgPPK_mCherry construct. The black arrow indicates where the expected band should be (66 kDa) in the elution fraction lane. Ladder: Precision Plus Protein™ (Biorad)


</br> 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