Difference between revisions of "Part:BBa K4323000"
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<partinfo>BBa_K4323000 short</partinfo> | <partinfo>BBa_K4323000 short</partinfo> | ||
| − | Secreted, cytolytic toxin that forms pores in host membranes after proteolytic removal of a C-terminal propeptide, leading to destruction of the membrane permeability barrier and host cell death. The pores are formed by transmembrane beta-strands and are approximately 3 nm in diameter. This part | + | Secreted, cytolytic toxin that forms pores in host membranes after proteolytic removal of a C-terminal propeptide, leading to destruction of the membrane permeability barrier and host cell death. The pores are formed by transmembrane beta-strands and are approximately 3 nm in diameter. This part is a coding sequence with a polyhistidine tag for purification. This part is codon optimized for expression in <i>Escherichia coli</i>. The composite part [https://parts.igem.org/Part:BBa_K4323001 BBa_K4323001] includes the necessary components to express Aerolysin toxin. For more information, please visit our [https://2022.igem.wiki/umanitoba/ Wiki page]. |
[[File:Aerolysin toxicity.png|800px|center|Aerolysin toxicity]] | [[File:Aerolysin toxicity.png|800px|center|Aerolysin toxicity]] | ||
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[[File:Ni-purified Aerolysin SDS-PAGE.png|400px|center|Ni-purified Aerolysin SDS-PAGE]] | [[File:Ni-purified Aerolysin SDS-PAGE.png|400px|center|Ni-purified Aerolysin SDS-PAGE]] | ||
| − | '''Figure 2. His-Trap Ni-column purified Aerolysin. L: ladder, C5: 10 % B wash, D1: fraction of gradient at ~30 % B, D4: fraction of gradient at ~50 % B, D8: elution at 100 % B. A buffer: 20 mM Tris-HCl, 200 mM NaCl, pH 8.0; B buffer: 20 mM Tris-HCl, 200 mM NaCl, 300 mM imidazole, pH 8.0. Arrow: purified Aerolysin protein, MW: 55.2 kDa.''' | + | '''Figure 2. His-Trap Ni-column purified Aerolysin. L: ladder, C5: 10 % B wash, D1: fraction of gradient at ~30 % B, D4: fraction of gradient at ~50 % B, D8: elution at 100 % B. A buffer: 20 mM Tris-HCl, 200 mM NaCl, pH 8.0; B buffer: 20 mM Tris-HCl, 200 mM NaCl, 300 mM imidazole, pH 8.0. Arrow: purified Aerolysin protein, MW: 55.2 kDa. 15 kDa and 10 kDa markers are not resolved.''' |
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Latest revision as of 20:42, 13 October 2022
Coding sequence for Aerolysin
Secreted, cytolytic toxin that forms pores in host membranes after proteolytic removal of a C-terminal propeptide, leading to destruction of the membrane permeability barrier and host cell death. The pores are formed by transmembrane beta-strands and are approximately 3 nm in diameter. This part is a coding sequence with a polyhistidine tag for purification. This part is codon optimized for expression in Escherichia coli. The composite part BBa_K4323001 includes the necessary components to express Aerolysin toxin. For more information, please visit our Wiki page.
Figure 1. The toxicity of Aerolysin to Dreissena bugensis.
Figure 2. His-Trap Ni-column purified Aerolysin. L: ladder, C5: 10 % B wash, D1: fraction of gradient at ~30 % B, D4: fraction of gradient at ~50 % B, D8: elution at 100 % B. A buffer: 20 mM Tris-HCl, 200 mM NaCl, pH 8.0; B buffer: 20 mM Tris-HCl, 200 mM NaCl, 300 mM imidazole, pH 8.0. Arrow: purified Aerolysin protein, MW: 55.2 kDa. 15 kDa and 10 kDa markers are not resolved.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 206
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 75
Illegal AgeI site found at 313
Illegal AgeI site found at 1270 - 1000COMPATIBLE WITH RFC[1000]