Difference between revisions of "Part:BBa K1985010"
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===Usage and Biology=== | ===Usage and Biology=== | ||
| − | This part is an improved version of a previously designed BioBrick (Part: | + | This part is an improved version of a previously designed BioBrick (Part:[https://parts.igem.org/Part:BBa_K1739000 BBa_K1739000]) from the Kent 2015 iGEM team. This part contains two segments, the CsgA signal sequence and only the first 61 aminoacids of the prion domain Sup35. Our improved BioBrick aims to optimize the self-assembly process of amyloid fibrils with the addition of these residues as they have been considered to be a suitable building block for the assembly of functional nanostructures. This improved plasmid does not contain the constitutive promoter (Part:[https://parts.igem.org/Part:BBa_J23104 BBa_J23104]) as was used for (Part:[https://parts.igem.org/Part:BBa_K1739000 BBa_K1739000]), giving the user choice over the promoter that they use. |
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===Validation=== | ===Validation=== | ||
| − | The plasmid was analysed through a diagnostic double restriction cut, using the enzymes NdeI and SspI. This was followed by agarose gel electrophoresis. The enzymes cleave the insert and a part of the plasmid at 1618bp, with the remainder plasmid being 790 bp. The sizes of the two fragments were compared with the size of the Invitrogen 1kB DNA marker and was found that our fragments were the correct | + | The plasmid was analysed through a diagnostic double restriction cut, using the enzymes NdeI and SspI. This was followed by agarose gel electrophoresis. The enzymes cleave the insert and a part of the plasmid at 1618bp, with the remainder plasmid being 790 bp. The sizes of the two fragments were compared with the size of the Invitrogen 1kB DNA marker and was found that our fragments were the correct sizes. |
[[File:PSB1C3+61.jpeg||400px|thumb|centre|Figure 1.1% agarose gel of the restriction digest of BBa_K1985010 in pSB1C3 plasmid backbone with NdeI and SspI]] | [[File:PSB1C3+61.jpeg||400px|thumb|centre|Figure 1.1% agarose gel of the restriction digest of BBa_K1985010 in pSB1C3 plasmid backbone with NdeI and SspI]] | ||
Revision as of 18:39, 19 October 2016
Sequence coding for amyloid Sup35 residues 1-61
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]
Usage and Biology
This part is an improved version of a previously designed BioBrick (Part:BBa_K1739000) from the Kent 2015 iGEM team. This part contains two segments, the CsgA signal sequence and only the first 61 aminoacids of the prion domain Sup35. Our improved BioBrick aims to optimize the self-assembly process of amyloid fibrils with the addition of these residues as they have been considered to be a suitable building block for the assembly of functional nanostructures. This improved plasmid does not contain the constitutive promoter (Part:BBa_J23104) as was used for (Part:BBa_K1739000), giving the user choice over the promoter that they use.
Validation
The plasmid was analysed through a diagnostic double restriction cut, using the enzymes NdeI and SspI. This was followed by agarose gel electrophoresis. The enzymes cleave the insert and a part of the plasmid at 1618bp, with the remainder plasmid being 790 bp. The sizes of the two fragments were compared with the size of the Invitrogen 1kB DNA marker and was found that our fragments were the correct sizes.
