Difference between revisions of "Part:BBa K1151000:Design"

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===Design Notes===
 
===Design Notes===
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It was inserted in a pET-15b plasmid vector, in order to have a better storage.
 
  
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This part was amplified by PCR from the plasmid received.
  
 +
                                [[File:nikRPCR.jpg]]
 +
 +
'''Figure 1:''' PCR results of HpNikR cds.
 +
 +
 +
'''Primers used''' (including Biobricks' Prefix and Suffix, lowercase):
 +
 +
''HpNikRFor:'' gtttcttcgaattcgcggccgcttctagATGGATACACCCAATAAAGACG
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''HpNikRRev:'' gtttcttcctgcagcggccgctactagtattattaCTATTCATTGTGTTCAAAG
  
 
===Source===
 
===Source===
  
−
It was amplified by PCR from the genomic sequence of Helicobacter pylori G27.  
+
We received this part by Dr. Alberto Danielli, a researcher in molecular biology at the Department of Pharmacology and Biotechnology at the University of Bologna. He sent us the gene encoding HpNikR (from the genome of ''H. pylori G27'') inserted into a pET-15b plasmid.
  
 
===References===
 
===References===
  
−
1.  Muller et al, Hierarchical regulation of the NikR-mediated nickel response in Helicobacter pylori. Nucleic Acids Res. 2011; 39(17):7564-75.
+
1.  Muller et al., Hierarchical regulation of the NikR-mediated nickel response in Helicobacter pylori. Nucleic Acids Res. 2011; 39(17):7564-75.
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2.  Dian  et al,  Structural Basis of the Nickel Response in Helicobacter pylori: Crystal Structures of HpNikR in Apo and Nickel -bound States.  J Mol Biol. 2006;  
+
 
−
25;361(4):715-30.  
+
2.  Dian  et al.,  Structural Basis of the Nickel Response in Helicobacter pylori: Crystal Structures of HpNikR in Apo and Nickel -bound States.  J Mol Biol. 2006; 25;361(4):715-30.  
 +
 
 
3.  Dosanjh et Michel, Microbial nickel metalloregulation: NikRs for nickel ions. Curr Opin Chem Biol. 2006 (2):123-30.
 
3.  Dosanjh et Michel, Microbial nickel metalloregulation: NikRs for nickel ions. Curr Opin Chem Biol. 2006 (2):123-30.
−
4.  Ernst  et al,  The nickel-responsive regulator NikR controls activation and repression of gene transcription in Helicobacter pylori .  Infect Immun.  2005;  
+
 
−
73(11):7252-8.
+
4.  Ernst  et al.,  The nickel-responsive regulator NikR controls activation and repression of gene transcription in Helicobacter pylori .  Infect Immun.  2005; 73(11):7252-8.
 +
 
 +
5. Abraham et al., The metal- and DNA-binding activities of Helicobacter pylori NikR. Journal of Inorganic Biochemistry. 100 (2006): 1005–1014.
 +
 
 +
6. West et al., Holo-Ni(II)HpNikR is an asymmetric tetramer containing two different nickel binding sites. J Am Chem Soc. 2010 October 20; 132(41): 14447–14456.

Latest revision as of 11:08, 3 October 2013

Nickel-responsive pleiotropic regulator (HpNikR)


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]


Design Notes

This part was amplified by PCR from the plasmid received. 

                               NikRPCR.jpg

Figure 1: PCR results of HpNikR cds.


Primers used (including Biobricks' Prefix and Suffix, lowercase):

HpNikRFor: gtttcttcgaattcgcggccgcttctagATGGATACACCCAATAAAGACG

HpNikRRev: gtttcttcctgcagcggccgctactagtattattaCTATTCATTGTGTTCAAAG

Source

We received this part by Dr. Alberto Danielli, a researcher in molecular biology at the Department of Pharmacology and Biotechnology at the University of Bologna. He sent us the gene encoding HpNikR (from the genome of H. pylori G27) inserted into a pET-15b plasmid.

References

1. Muller et al., Hierarchical regulation of the NikR-mediated nickel response in Helicobacter pylori. Nucleic Acids Res. 2011; 39(17):7564-75.

2. Dian et al., Structural Basis of the Nickel Response in Helicobacter pylori: Crystal Structures of HpNikR in Apo and Nickel -bound States. J Mol Biol. 2006; 25;361(4):715-30.

3. Dosanjh et Michel, Microbial nickel metalloregulation: NikRs for nickel ions. Curr Opin Chem Biol. 2006 (2):123-30.

4. Ernst et al., The nickel-responsive regulator NikR controls activation and repression of gene transcription in Helicobacter pylori . Infect Immun. 2005; 73(11):7252-8.

5. Abraham et al., The metal- and DNA-binding activities of Helicobacter pylori NikR. Journal of Inorganic Biochemistry. 100 (2006): 1005–1014.

6. West et al., Holo-Ni(II)HpNikR is an asymmetric tetramer containing two different nickel binding sites. J Am Chem Soc. 2010 October 20; 132(41): 14447–14456.