Team:TU-Eindhoven/Basic Part
Basic Parts
The list below shows our Basic Parts. For the medal requirements, we nominated BBa_K2812000 and BBa_K2812004 for the silver and gold medal, respectively. As new part for the silver medal, we created the carbohydrate binding domain, BBa_K2812000, an adhesin binding strongly to glucose molecules. For the gold medal, TU-Eindhoven 2018 functionally improved on the original truncated lysostaphin BBa_K748002 created by HIT-Harbin 2012 team in multiple ways. Through codon optimization, expression of the lysostaphin domain has been improved considerably. The other major functional improvement was realized by fusing the truncated lysostaphin to the HlyA secretion domain via a cleavable thrombin linker. In this way, (continuous) secretion via the Type I pathway can be achieved by co-expressing it with the HlyB/D secretion proteins. This avoids the requirement of cell lysis for release of the truncated lysostaphin, improving on the project of iGEM Harbin 2012, as self-destruction of their killer device is no longer necessary. Additionally, this new construct simplifies purification of truncated lysostaphin as running the medium over a nickel affinity column followed by thrombin cleavage results in pure truncated lysostaphin. The part BBa_K2812004 has been characterized extensively and the expression, secretion and selective activity of the truncated lysostaphin against S. aureus is demonstrated successfully.
| Fave Part | Name | Type | Description | Designers | Length |
|---|---|---|---|---|---|
| ❤ | BBa_K2812004 | Basic | Coding sequence for trunctated Lysostaphin fused to His-tagged HlyA | Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls | 1458 |
| ❤ | BBa_K2812000 | Basic | Carbohydrate-binding domain from MpIBP | Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls | 576 |
| BBa_K2812001 | Basic | Coding sequence for trunctated Lysostaphin | Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls | 738 | |
| BBa_K2812002 | Basic | pBAD-ara promoter - Arabinose inducible regulatory promoter | Guido Oerlemans, Maxime van den Oetelaar and Mariska Brüls | 71 | |
| BBa_K2812006 | Basic | Coding sequence for Pyocin S5 with HlyA and His6-tag | Guido Oerlemans, Maxime van den Oetelaar & Mariska Brüls | 2217 |
