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Konstruktion av reporterplasmider innehållande möjliga promotorregioner för kloratreduktas respektive kloritdismutas från Ideonella dechloratans
Karlstad University, Faculty of Technology and Science, Department of Chemistry and Biomedical Sciences.
2012 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
Construction of reporter plasmids containing possible promoter regions of chlorate reductase and chlorite dismutase from Ideonella dechloratans (English)
Abstract [sv]

Ideonella dechloratans är en av flera isolerade bakteriestammar med förmågan att använda klorat i sin metabolism i anaerob miljö. Detta kan utnyttjas inom exempelvis pappersindustrin där klorat är en miljöfarlig restprodukt efter klordioxidblekning.

   Klorat och perklorat har visat sig ha negativa effekter hos både människor, djur och växter vilket ger ett stort behov av mer forskning på förbättrade reningsmetoder. Kan genregleringen av enzymerna som sköter nedbrytningen av klorat, kloratreduktas och kloritdismutas, förbättras genom att exempelvis fungera även i aerob miljö så skulle reningen förbättras samt kostnader dras ner.

   Genom att införa promotorregionerna för kloratreduktas och kloritdismutas i en broad-host-range reporterplasmid, pQlacZ-1, så kan dessa testas i flera olika förhållanden och i olika typer av gramnegativa bakterier.

   Genom dubbelklyvning av pQlacZ-1 med BamHI och EcoRI så har PCR produkter av Clrp och Cldp från Ideonella dechloratans kunnat ligeras in för att sedan transformeras in i E. coli, XL-1 Blue.    Gelextraktion har visat sig vara den effektivaste reningsmetoden inför ligering men utförligare screening behöver göras på transformanterna för att säkerställa metodens effektivitet.

Abstract [en]

Ideonella dechloratans is one of several isolated strains of bacteria with the ability to use chlorate in its metabolism in an anaerobic environment. This can be utilized by, for example, the paper industry where chlorate is an environmentally dangerous pollutant after chlorine dioxine bleaching. 

   Chlorate and perchlorate have been found to have negative effects in humans, animals and plants which give rise to a need for more research on improved purification methods. If the gene regulation of the enzymes that handle the breakdown of chlorate, chlorate reductase and chlorite dismutase, could be improved to work in an aerobic environment then both the purification and cost efficiency could improve.

   By inserting the promoter regions for chlorate reductase and chlorite dismutase in a broad-host-range reporterplasmid, pQlacZ-1, they can be evaluated in several different conditions and different strains of gram-negative bacteria.

   Through double cleaving of pQlacZ-1 with BamHI and EcoRI, PCR products of Clrp and Cldp from Ideonella dechloratans have been ligated into E. coli, XL-1 Blue.

   Gel extraction was found to be the most efficient method of purification before ligation but further screening needs to be performed on the transformants to ensure the efficiency of the method.

Place, publisher, year, edition, pages
2012. , p. 26
Keywords [sv]
Ideonella dechloratans, pQlacZ-1, kloritdismutas, kloratreduktas, promotor
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kau:diva-13720Local ID: KEM-31OAI: oai:DiVA.org:kau-13720DiVA, id: diva2:534084
Subject / course
Chemistry
Uppsok
Physics, Chemistry, Mathematics
Supervisors
Examiners
Available from: 2012-06-26 Created: 2012-06-15 Last updated: 2012-06-26Bibliographically approved

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