Difference between revisions of "Part:BBa K1151000"

(Cytosol/membrane separation by Zerial method)
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Figure 4: PAGE of the separation.
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'''Figure 4:''' PAGE of the separation.
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 13:16, 26 September 2013

Nickel-responsive pleiotropic regulator (HpNikR)

The HpNikR protein is a pleiotropic regulator from Helicobacter pylori. In presence of nickel it can acts as an activator or a repressor depending of the specific promoter that contains its operator site. It consists of two dimeric DNA binding domains separated by a tetrameric regulatory domain that binds nickel. This domain corresponds to the C-terminal regulatory domain which contains four nickel binding sites at the tetramer interface. Binding nickel, then a conformational change allows it to activate or repress trascription.


Usage and Biology

                                       Cattura.jpg                                           

Figure 1: HpNikR nickel-binding domain (tetramer).



Cattura1.jpg

Figure 2: Proteic sequence analysis with Uniprot.


NikR expression using BL21 (DE3) cells

First we made ​​competent BL21 cells and we transformed it with the plasmid containing NikR. We then proceeded with the normal protocol of induction with IPTG.

Foto1.jpg

Figure 3: PAGE of the first induction.


Cytosol/membrane separation by Zerial method

To confirm that NikR is a cytosolic protein (not expressed in multivesicular bodies, and then in membrane) we performed a separation membrane-cytosol (Zerial method.

Foto2.jpg

Figure 4: PAGE of the separation.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]