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Construction of a fusion protein for anchoring the inflammatory receptor NLRP3 to the cell membrane
University of Skövde, School of Bioscience.
2019 (English)Independent thesis Basic level (degree of Bachelor), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

The innate immune system are a cooperation of many components – receptors being one of them. Both membrane-bound and cytosolic receptors play a large role in the defence system against pathogens and danger. NLRP3 is a receptor which assembles a protein complex called inflammasome in response to cytosolic stress and is responsible for many autoimmune diseases if it malfunctions. The activation of the NLRP3 inflammasome leads to secretion of inflammatory cytokines and in many cases to programmed cell death. The structure, function and activation of the NLRP3 inflammasome is still not fully understood and the urge to understand the mechanisms behind are important for future medical improvements. The aim was to anchor the NLRP3 inflammasome by the cell membrane - By Overlap PCR, the NLRP3 cDNA was fused extracellular and trans-membrane parts of the TLR4 cDNA to anchor the NLRP3 to the membrane and in turn analyse the inflammasome with LPI™ technology. Multiple primers and a TLR4 nucleotide were designed and the NLRP3 was amplified with specific overhangs by PCR. The fusion protein was successfully linked together by Overlap PCR but not confirmed by sequencing. The gene fusion demands high quality primers for amplification and further evaluation must be made to the details of the laboratory. To anchor the protein complex to the cell membrane, continue to be of full importance and can be an asset in many structural studies and biopharmaceuticals trials.

Place, publisher, year, edition, pages
2019. , p. 26
Keywords [en]
NLRP3 inflammasome, Overlap PCR, Fusion Protein, LPI™ technology
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:his:diva-17482OAI: oai:DiVA.org:his-17482DiVA, id: diva2:1339167
Subject / course
Bioscience
Educational program
Bioscience - Molecular Biodesign
Supervisors
Examiners
Available from: 2019-08-08 Created: 2019-07-26 Last updated: 2019-09-10Bibliographically approved

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CiteExportLink to record
Permanent link

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Citation style
  • apa
  • ieee
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  • vancouver
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  • de-DE
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  • en-US
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  • nn-NB
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  • Other locale
More languages
Output format
  • html
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