Expression and purification of plant proteins for functional studies
Independent thesis Advanced level (degree of Master (One Year)), 30 credits / 45 HE creditsStudent thesis
Chloroplasts are cell organelles responsible for photosynthesis. Although chloroplast have their own genome it is not sufficient to encode all the proteins which are located there. Most of the proteins are imported from the cytosol through the so called toc/tic pathway. It has been recently showed that Arabidopsis CAH1 is transported to the chloroplast through the secretory route in a fully new pathway. It has also been demonstrated that the N-terminal signal peptide of CAH1 targets it to the ER where the protein gets glycosylated. Structure of the Arabidopsis CAH1 suggests that its C-terminus might be responsible for targeting the protein to the chloroplast. By expressing N- and C-terminal labeled CAH1 we show that the expression level of the N-terminal labeled form is high and the majority of the labeled protein is localized in the chloroplast. By contrast, the C-terminal labeled CAH1 expressed weakly if at all, and due to the low expression level immunolocalization of the protein is difficult. We also demonstrate that the strong expression level of the N-terminal labeled CAH1 makes it feasible to affinity purify the glycosylated protein for structural studies.
Place, publisher, year, edition, pages
2006. , 13 p.
IdentifiersURN: urn:nbn:se:umu:diva-26082OAI: oai:DiVA.org:umu-26082DiVA: diva2:236495
KBF30, UPSC, Umeå universitet, Umeå (English)
Bako, Laszlo, Univ lekt
Jansson, Stefan, Prof