Difference between revisions of "Part:BBa K1679029"

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===Usage and Biology===
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===Experiment===
[[File:ftnA_native page.jpg|px1000|thumb|left|Figure 2. Native PAGE analysis of mineralized FtnA
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We purified the ferritin overexpressed in E.coli through Ni-chelating affinity chromatography and highly concentrated it and then do SDS-PAGE. The position of clearly targeted band(19.5kDA) on the gel was consistent with the size of ferritin monomer fused with tag on plasmid, explaining that the ftnA expression is successful.
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[[File:ferritin concentrated purified product SDS-PAGE.jpg|px1000|thumb|left|Figure 2. SDS-PAGE shows the expected protein band.]]
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[[File:ftnA_native page.jpg|px1000|thumb|left|Figure 3. Native PAGE analysis of mineralized FtnA
 
Gel was stained with (A) potassium ferrocyanide and (B) Coomassie Brilliant Blue R250. Control, HFn; lane 1, concentrated ferritin which purified and concentrated from group 1; lane 2, bacterium sediment of group 1; lane 3, concentrated ferritin which purified and concentrated from group 3; lane 4, bacterium sediment of group 3.]]
 
Gel was stained with (A) potassium ferrocyanide and (B) Coomassie Brilliant Blue R250. Control, HFn; lane 1, concentrated ferritin which purified and concentrated from group 1; lane 2, bacterium sediment of group 1; lane 3, concentrated ferritin which purified and concentrated from group 3; lane 4, bacterium sediment of group 3.]]
  

Revision as of 18:33, 18 September 2015

ftnA

FtnA is a bacterial ferritin with a protein shell is assembled from 24 identical 19.4 kDa FtnA monomers. Its central cavity is around 7.5 nm in diameter and can be loaded with iron when cells grow under iron-rich conditions[1]. The iron is stored in the form of ferrihydrite iron cores normally that with superparamagnetic properties[2]. The iron contained ferritin can generate heat in response to electromagnetic signal[3]. For the reasons above, we design it as our magnetic receiver which can turn electromagnetic signal into heat.

[1]Smith J L. The physiological role of ferritin-like compounds in bacteria[J]. Critical reviews in microbiology, 2004, 30(3): 173-185. [2] Papaefthymiou G C, Viescas A J, Devlin E, et al. Electronic and magnetic characterization of in vivo produced vs. in vitro reconstituted horse spleen ferritin[C]//MRS Proceedings. Cambridge University Press, 2007, 1056: 1056-HH03-27. [3] Stanley S A, Sauer J, Kane R S, et al. Remote regulation of glucose homeostasis in mice using genetically encoded nanoparticles[J]. Nature medicine, 2015, 21(1): 92-98.

Fig.1. Schema of ferritin


Experiment

We purified the ferritin overexpressed in E.coli through Ni-chelating affinity chromatography and highly concentrated it and then do SDS-PAGE. The position of clearly targeted band(19.5kDA) on the gel was consistent with the size of ferritin monomer fused with tag on plasmid, explaining that the ftnA expression is successful.

Figure 2. SDS-PAGE shows the expected protein band.
Figure 3. Native PAGE analysis of mineralized FtnA Gel was stained with (A) potassium ferrocyanide and (B) Coomassie Brilliant Blue R250. Control, HFn; lane 1, concentrated ferritin which purified and concentrated from group 1; lane 2, bacterium sediment of group 1; lane 3, concentrated ferritin which purified and concentrated from group 3; lane 4, bacterium sediment of group 3.


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]

Functional Parameters