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Team:Washington/Project
From 2009.igem.org
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== Project Details== | == Project Details== | ||
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| + | The target system is a biobrick compatible expression vector, that tags any favorite protein to be secreted, purified, and localized on the cell surface. To do this, we synthesized our “target construct” through gene synthesis to have a nano-tag, two histidine tags, two TEV protease sights, an Nhe1 cut site, and prtB (need biobrick name) and terminator BBa_B0015(all need referenecs). The Nhe1 sticky ends are compatible with Xba1 and Spe1 sticky ends, allowing for the insertion of biobricked genes into the target construct. The nano-tag is a short sequence that allows the target construct to non-covalently bond to the surface protein streptavadin, in a highly characterized interaction (reference). The Harvard part ?????? from 2006, a monomeric form of streptavadin theoretically allows the protein to linger on the cell till a convenient time for purification, though we had difficulty getting functional protein (please reference display team). TEV protease sights flank the Nhe1 sight allowing the desired protein to be excised from the target construct if desired during or after purification. Histidine tags are on the outsides of the TEV sights allowing the target construct with a favorite protein to be purified via the traditional purification methods, or to remove fragments of the target construct after TEV cutting, leaving highly purified protein behind. | ||
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Revision as of 20:17, 6 October 2009
| Secretion System | Display System |
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ENTER IN PROJECT BACKGROUND INFO HERE!
Original Page, get rid of later---------
| You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing. |  |
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Tell us more about your project. Give us background. Use this is the abstract of your project. Be descriptive but concise (1-2 paragraphs) |  |
| Team Example 2 |
| Home | The Team | The Project | Parts Submitted to the Registry | Modeling | Notebook |
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(Or you can choose different headings. But you must have a team page, a project page, and a notebook page.)
Overall project
Your abstract
Project Details
The target system is a biobrick compatible expression vector, that tags any favorite protein to be secreted, purified, and localized on the cell surface. To do this, we synthesized our “target construct” through gene synthesis to have a nano-tag, two histidine tags, two TEV protease sights, an Nhe1 cut site, and prtB (need biobrick name) and terminator BBa_B0015(all need referenecs). The Nhe1 sticky ends are compatible with Xba1 and Spe1 sticky ends, allowing for the insertion of biobricked genes into the target construct. The nano-tag is a short sequence that allows the target construct to non-covalently bond to the surface protein streptavadin, in a highly characterized interaction (reference). The Harvard part ?????? from 2006, a monomeric form of streptavadin theoretically allows the protein to linger on the cell till a convenient time for purification, though we had difficulty getting functional protein (please reference display team). TEV protease sights flank the Nhe1 sight allowing the desired protein to be excised from the target construct if desired during or after purification. Histidine tags are on the outsides of the TEV sights allowing the target construct with a favorite protein to be purified via the traditional purification methods, or to remove fragments of the target construct after TEV cutting, leaving highly purified protein behind.
