Development of abscisic acid biosensors in yeast

Projektförslag för kandidatarbete inom inst. Kemi och kemiteknik och Biologi och bioteknik

Projektexamenskod: BBTX01-20-02

Avdelningen för Systembiologi
Institutionen för Biologi och Bioteknik, Chalmers tekniska högskola

Bakgrund
Yeast has become one of the preferred cell factories for the sustainable production of fuels and chemicals. In our group, we recently constructed a yeast strain able to produce the plant hormone abscisic acid (ABA), which plays a major role in stress resistance in plants but also has potential medical applications. However, the production levels of this engineered yeast strain are still low most likely due to low enzyme activity levels. The availability of suitable ABA biosensors would make it possible to screen enzyme libraries for increased production levels.

Problembeskrivning
The aim of this project is to construct and test a range of different ABA biosensors in the yeast Saccharomyces cerevisiae. These will be based on two plant proteins that interact only on the presence of ABA. To connect this protein-protein-interaction with a visible output signal (expression of a fluorescent protein) a yeast-to-hybrid and a CRISPRi-based system will be tested.

Genomförande /Viktiga moment/teknikinnehåll
1. Project preparation / literature studies
    a. Study of provided papers and additional literature search
    b. Study of previously conducted experiments
    c. Design of test system(s) and controls
    d. PCR primer design
2. Implementation
    a. PCR amplification of elements
    b. Plasmid assembly and yeast transformation
3. Evaluation
    a. Sensor evaluation under different conditions and/or in different background strains using flow cytometry

Speciella förkunskapskrav: Basic knowledge in cell and molecular biology is required.

Möjlig målgrupp: Preferably ”Bioteknik” students, but open for students from other programmes, if they have the required background knowledge.

Gruppstorlek: 4–6 studenter

Förslagsställare/kontaktperson/huvudhandledare: Verena Siewers (siewers@chalmers.se)
Övriga handledare: Florian David, Maximilian Otto, Yasaman Dabirian
Ev. förslag på examinator: Nathalie Scheers

Publicerad: ti 29 okt 2019.