Difference between revisions of "Part:BBa K1739001"
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<partinfo>BBa_K1739001 short</partinfo>
 
<partinfo>BBa_K1739001 short</partinfo>
  
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This part uses the BBa_J23104 and encodes cytochrome b562 in a pSB1C3 backbone. This part has been validated by digestion and quantification of the presence of the cytochrome gene on a diagnostic gel. Cytochrome b562 is a single subunit, four-helix bundle protein containing a non-covalently bound b-type haem group with a molecular weight of 25kDa (Fujiwara, Fnkumori, and Yamanaka, 1993; Robinson et al., 1997).  
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This part uses the BBa_J23104 and encodes Cytochrome <i>b</i><sub>562</sub> in a pSB1C3 backbone. This part has been validated by digestion and quantification of the presence of the cytochrome gene on a diagnostic gel. Cytochrome <i>b</i><sub>562</sub> is a single subunit, four-helix bundle protein containing a non-covalently bound b-type haem group with a molecular weight of 25kDa (Fujiwara, Fnkumori, and Yamanaka, 1993; Robinson et al., 1997).  
  
 
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===Validation===
 
===Validation===
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To validate the nature of our part, we analysed it through agarose gel electrophoresis. A restriction digest preceded this, using the enzymes ECORI and PSTI. The enzymes cleave pSBIC3 into a fragment of 2030bp. By comparing the sizes of the fragments to a marker we discovered the size of our insert ( as seen in figure 1) to be around 400bp, matching the actual size of 364 base pairs of the Cyt<i>b</i><sub>562</sub> sequence. Also, our gel shows a fragment to be about 2000 base pairs, confirming it is pSB1C3 which is 2030 base pairs in size.
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[[File:Team Kent Cytb562gel labelled.jpg|thumb|center|400px|Figure 1. Shows the agarose gel, following a restrictive digest of our BioBrick containing Cytochrome <i>b</i><sub>562</sub>.]]
  
  

Revision as of 16:08, 18 September 2015

Sequence coding for Cytochrome b562

This part uses the BBa_J23104 and encodes Cytochrome b562 in a pSB1C3 backbone. This part has been validated by digestion and quantification of the presence of the cytochrome gene on a diagnostic gel. Cytochrome b562 is a single subunit, four-helix bundle protein containing a non-covalently bound b-type haem group with a molecular weight of 25kDa (Fujiwara, Fnkumori, and Yamanaka, 1993; Robinson et al., 1997).

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]


Validation

To validate the nature of our part, we analysed it through agarose gel electrophoresis. A restriction digest preceded this, using the enzymes ECORI and PSTI. The enzymes cleave pSBIC3 into a fragment of 2030bp. By comparing the sizes of the fragments to a marker we discovered the size of our insert ( as seen in figure 1) to be around 400bp, matching the actual size of 364 base pairs of the Cytb562 sequence. Also, our gel shows a fragment to be about 2000 base pairs, confirming it is pSB1C3 which is 2030 base pairs in size.

Figure 1. Shows the agarose gel, following a restrictive digest of our BioBrick containing Cytochrome b562.


References:

Fujiwara, T., Fnkumori,, Y. and Yamanaka, T. (1993). Halobacterium halobium Cytochrome b-558 and Cytochrome b-562: Purification and Some Properties. J. Biochem., 113, pp.48-54.

Robinson, C., Liu, Y., Thomson, J., Sturtevant, J. and Sligar, S. (1997). Energetics of Heme Binding to Native and Denatured States of Cytochrome b 562 †. Biochemistry, 36(51), pp.16141-16146.