Difference between revisions of "Part:BBa K3739022"
Line 3: | Line 3: | ||
<partinfo>BBa_K3739022 short</partinfo> | <partinfo>BBa_K3739022 short</partinfo> | ||
− | Ag43 is a surface display protein and GFP can show the expression effciency of Ag43 | + | Ag43 is a surface display protein and GFP can show the expression effciency of Ag43. |
− | + | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
− | Ag43 is a surface display protein that displays | + | Ag43 is a surface display protein that displays protein molecule it carries on the cell membrane of bacteria. pelB is an exopeptide linked to Ag43. Tevp can shear Ag43 and release the exopeptide pelB to secrete the protein molecules out of bacteria. |
+ | |||
+ | We used <partinfo>BBa_K525998</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K3739113</partinfo>, which is assembled on the expression vector pET-28(+) by standard assembly. The constructed plasmid was transformed into ''Vibrio natriegens'' , then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing. | ||
===Reference=== | ===Reference=== |
Revision as of 04:57, 21 October 2021
pelB-Ag43-160N-his-GFP
Ag43 is a surface display protein and GFP can show the expression effciency of Ag43.
Usage and Biology
Ag43 is a surface display protein that displays protein molecule it carries on the cell membrane of bacteria. pelB is an exopeptide linked to Ag43. Tevp can shear Ag43 and release the exopeptide pelB to secrete the protein molecules out of bacteria.
We used BBa_K525998 to construct the expression system and obtained the composite BBa_K3739113, which is assembled on the expression vector pET-28(+) by standard assembly. The constructed plasmid was transformed into Vibrio natriegens , then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing.
Reference
1. Ahan, R. E.; Kirpat, B. M.; Saltepe, B.; Seker, U. O. S., A Self-Actuated Cellular Protein Delivery Machine. ACS Synth Biol 2019, 8 (4), 686-696.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 54
Illegal NgoMIV site found at 1012
Illegal NgoMIV site found at 2236
Illegal NgoMIV site found at 2746
Illegal NgoMIV site found at 2767
Illegal AgeI site found at 1762
Illegal AgeI site found at 2506
Illegal AgeI site found at 2920
Illegal AgeI site found at 3162 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 142