Difference between revisions of "Part:BBa K1151009"
 
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This is a nickel sensitive system driven by the promoter pnikR ([[BBa_K1151003]]) with a GFP reporter. Normally the production of GFP is switched on, it is possible to repress the GFP expression using nickel ions in medium and in presence of HpNikR ([[BBa_K1151000]]) as trascription factor.
 
This is a nickel sensitive system driven by the promoter pnikR ([[BBa_K1151003]]) with a GFP reporter. Normally the production of GFP is switched on, it is possible to repress the GFP expression using nickel ions in medium and in presence of HpNikR ([[BBa_K1151000]]) as trascription factor.
  
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===Boiling prep and digestion with restriction enzymes===
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===Usage and Biology===
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Once inserted the construct in a PSB1C3 plasmid (digestion and ligation), we transformed DH5a cells and then we recovered the ligated plasmid by boiling prep. We then proceeded with the digestion with EcoRI and PstI to check.
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                                [[File:EPUnisalento.jpg]]
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'''Figure 1:''' Digestion result of boiling prep plasmids.
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===PCR with VF2 and VR2 primers===
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                          [[File:VF23Unisalento.jpg]]
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'''Figure 2:''' PCR results.
  
 
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===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K1151009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1151009 SequenceAndFeatures</partinfo>

Latest revision as of 14:50, 2 October 2013

GFP nickel regulated 1

This is a nickel sensitive system driven by the promoter pnikR (BBa_K1151003) with a GFP reporter. Normally the production of GFP is switched on, it is possible to repress the GFP expression using nickel ions in medium and in presence of HpNikR (BBa_K1151000) as trascription factor.

Boiling prep and digestion with restriction enzymes

Once inserted the construct in a PSB1C3 plasmid (digestion and ligation), we transformed DH5a cells and then we recovered the ligated plasmid by boiling prep. We then proceeded with the digestion with EcoRI and PstI to check. 

                                EPUnisalento.jpg

Figure 1: Digestion result of boiling prep plasmids.

PCR with VF2 and VR2 primers

                          VF23Unisalento.jpg

Figure 2: PCR results.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 798