Difference between revisions of "Part:BBa K1151001"
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===PCR with VF2 and VR2 primers=== | ===PCR with VF2 and VR2 primers=== | ||
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'''Figure 2:''' PCR results. | '''Figure 2:''' PCR results. | ||
Revision as of 10:52, 2 October 2013
Histidine-rich metal-binding protein
So called for emphasize its origins in Helicobacter pylori and its avidity for nickel. Consisting of 60 aminoacids, the protein is rich in histidine (28 residues, 46.7 %) and contains short repeating motifs, it exists as an equilibration of multimeric forms in solution, with 20-mers (approx. 136 kDa) being the predominant species. Hpn can bind tightly and reversibly up to five Ni2+ ions per each monomer of 7 kDa in a pH-dependent manner (pH 7.4 ). In H. pylori play an important role in storing nickel required to the survival of the bacterium.
Boiling prep and digestion with restriction enzymes
Once inserted the gene coding for Hpn in a PSB1C3 plasmid (digestion and ligation), we transformed DH5a cells with the construct and then we recovered the ligated plasmid by boiling prep. We then proceeded with the digestion with EcoRI and PstI to check.
Figure 1: Digestion result of boiling prep plasmids.
PCR with VF2 and VR2 primers
Figure 2: PCR results.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]