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Purification and analysis of Growth Transforming factor beta 1 expressed in plants


Faculty of Biology, Medicine and Health

About the Project

Recombinant human Transforming Growth Factor ß1 (hTGFß1) is in demand for use in R&D and to develop therapeutic agents for regenerative medicine and immunosuppressive therapy. The protein is involved in complex signalling pathways in which its location, timing and activity are important for its function. Two barriers limit applications of rhTGFß proteins. First, they are difficult and costly to manufacture by conventional platforms (yeast/bacteria/animal cells), which limits supply, and second, their use requires precise control of their activities in time and space to eliminate unintended effects.

This project focuses on the expression of hTGFbeta1 in plants. Plants provide a lean carbon expression platform that is highly scalable to express recombinant proteins. The expression is particularly high in the chloroplasts of plants. This allows large amounts of the protein to be expressed for functional characterisation in animal cell bioassays. It will also examine the potential of using chemical modification in vitro to study and control the activity of the protein.

The project will involve chloroplast expression of hTGFbeta1 to isolate the active homodimeric protein. The recombinant proteins will be purified from plants and refolded in vitro (Day).
The functional activity of the protein with and without chemical modification will be studied in animal cells (Travis).

Candidates are expected to hold (or be about to obtain) a minimum upper second class honours degree (or equivalent) in Biochemistry, Chemistry, Molecular Biology or a related subject Applicants with experience in protein chemistry or protein purification are encouraged to apply.

For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (https://www.bmh.manchester.ac.uk/study/research/apply/). Informal enquiries may be made directly to the primary supervisor. On the online application form select PhD Biochemistry.

For international students we also offer a unique 4 year PhD programme that gives you the opportunity to undertake an accredited Teaching Certificate whilst carrying out an independent research project across a range of biological, medical and health sciences. For more information please visit http://www.internationalphd.manchester.ac.uk





Funding Notes

Applications are invited from self-funded students. This project has a Band 3 fee. Details of our different fee bands can be found on our website (View Website). For information on how to apply for this project, please visit the Faculty of Biology, Medicine and Health Doctoral Academy website (View Website).

As an equal opportunities institution we welcome applicants from all sections of the community regardless of gender, ethnicity, disability, sexual orientation and transgender status. All appointments are made on merit.

References

Gisby MF, Mellors P, Madesis P, Ellin M, Laverty H, O’Kane S, Ferguson MWJ, Day A (2011) A synthetic gene increases
TGFβ3 accumulation by 75-fold in tobacco chloroplasts enabling rapid purification and folding into a biologically active
molecule. Plant Biotechnology Journal 9:618-628.

Pagel, M. (2019) Inverse electron demand Diels-Alder (IEDDA) reactions in peptide chemistry. Journal of Peptide Science,
25. https://doi.org/10.1002/psc.3141

Kelly A , Houston SA, Sherwood E , Casulli J, Travis MA (2017). Regulation of Innate and Adaptive Immunity by TGFβ.
Adv Immunol., 134:137-233.

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