DEVMICRO · Directed Evolution in Femtolitre Microreactors - Exploring 'Promiscuity Space'
7РП — „Хора“ (Действия „Мария Кюри“)
- Период
- 2008-03-01 → 2010-02-28
- Финансиране от ЕС
- 177 173 €
- Участници
- 1
- Схема
- MC-IEF
Линиите свързват координатора с партньорите.
Накратко на български
Микрореакторите от водни капки в масло се тестват за изтичане на вещества, като например флуоресцентни молекули. Това помага да се намали взаимното замърсяване между отделните капки, което е важно за точното проследяване на химични реакции в единични клетки.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Directed Evolution in Femtolitre Microreactors - Exploring 'Promiscuity Space'
The use of water-inoil droplets as microreactors to carry out chemical and biochemical experiments relies on the assumption that the contents of droplets are not leaking out into the oil phase and there is no communication between the reaction vessels. To assess the mechanism of possible leaking, the retention of various fluorescein derivatives from droplets formed in mineral oil and stored for hours in a reservoir on chip was studied. Leaking into the oil phase was observed and was shown to be dependent on the charge of the compounds and on the concentration of the surfactant used. We found that the addition of bovine serum albumin substantially reduced leaking. The improved retention of small fluorescent molecules inside droplets on chip using our modified formulation was demonstrated by single-cell experiments in which encapsulated E. coli cells producing an enzyme were distinguished clearly from those without cells, suggesting that cross contamination of product was minimized.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
The ‘vastness of sequence space’, i.e. the extremely large number of possible permutations in even the smallest catalytic proteins, requires evolutionary shortcuts. In this project the candidate will interconvert members of a promiscuous catalytic superfamily into one anotehr and differentiate a multi-specific enzyme into mono-specific mutants. Ultimately it is hoped that directed evolution experiments will have implications for the definition of ‘phylogenetic relationships’ for catalysis. To this end the candidate will set up biochemical assays for hydrolytic reactions in small emulsion droplets (diameter: 2-20 um) and explore whether (i) these assays can be carried out in such in vitro compartemnts , (ii) whether it is possible to measure kinetics in these droplets oand (iii) carry out directed evolutino experiments with a sreening system of unprecedented capacity. In addition to tehse technical aspects, the candidate will also charcterise the mechanism of the wild type enzyme and its mutants.
Оригинален текст от CORDIS (на английски).
Участници
- THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE · CAMBRIDGEКоординаторОбединеното кралство
Връзки
Данни: CORDIS, © Европейски съюз