Difference between revisions of "Part:BBa K1489003"
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<partinfo>BBa_K1489003 short</partinfo> | <partinfo>BBa_K1489003 short</partinfo> | ||
| − | Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while | + | Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while an unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins. |
The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way. | The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way. | ||
Latest revision as of 04:49, 17 October 2014
Lpp-OmpA-Linker with KasI site
Outer membrane protein A (OmpA) is a common outer membrane protein found in bacteria that serves various purposes. For our purposes, the beta-barrel transmembrane domain can be utilized to anchor otherwise soluble proteins in the outer cell membrane. Lpp serves to direct the fusion protein, while an unstructured linker (GGGSGGGS) serves to separate OmpA from its cargo in order to avoid interactions between the two proteins.
The expressed protein is almost structurally identical to BBa_K103006, with the terminal serine on the linker replaced by an alanine. This change was made in order to convert the previous SacI site (GAGCTC), which was unusable for RE cloning due to a second SacI site being present in the plasmid backbone, into a KasI site (GGCGCC). The KasI site codes for Gly-Ala, while the SacI site codes for Gly-Ser. This amino acid substitution is unlikely to impact the function of Lpp-OmpA-Linker in a major way.
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]