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Cloning chitinase genes from Plasmodiophora brassicae into Escherichia coli
2017 (English)Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
Abstract [en]

Plasmodiophora brassicae is a soil-borne mono-cellular organism and a parasite on plants from the Brassicaceae family, which includes commercially important species such as cabbages, turnips, and rapeseed. The parasite infects the plant roots, causing swelling into galls which lead to decreased nutrition uptake and increased sensitivity to further infection from other microorganisms. These symptoms cause stunted growth and decreased yield from the infected plant.The organism’s spores are protected in the soil by a resilient, chitin-containing, spore wall. This wall let the spore survive for many years in the ground, and complicates genetic research on the organism as the wall makes DNA- and RNA-extraction more difficult.During this project, attempts were made to express genes for chitinases from P. brassicae in Escherichia coli, to see if the enzymes could be used to weaken the spore wall enough to facilitate further research. For this end, methods for RNA extraction, followed by PCR, cloning, and chitinase activity testing were used.Complete enzymes were not expressed during the timeframe of the project, but one of the genes, and part of a second one, were cloned and may be used in further attempts to properly express the encoded enzymes.

Place, publisher, year, edition, pages
2017.
Keywords [en]
Clubroot, spore wall, heterologous gene expression, plant parasites, resting spores
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:uu:diva-349211OAI: oai:DiVA.org:uu-349211DiVA, id: diva2:1199931
Educational program
Biomedical Laboratory Science Programme; Biomedical Laboratory Science Programme
Examiners
Available from: 2018-04-23 Created: 2018-04-23 Last updated: 2018-04-23Bibliographically approved

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