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Isolaion and characteization of an arsC generesponsible (gene ID: 2889) for arsenicsequestration in Lysinibacillus sphaericus
University of Skövde, School of Bioscience.
2018 (English)Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Many inhabited regions in the world, especially in the South‐East Asia, are suffering from heavy metalpollution in the soil, the ground water and in the corps. During the previous decades chemical methodswere being used to collect the pollutants from the environment, especially the arsenic, but during the lastyears bioremediation has gained ground, using microorganisms. Lysinibacillus sphaericus is one of thenumerous bacterial species that can be used for this purpose. It is a Gram+ bacterium that contains all theimportant genes for arsenic uptake and accumulation inside the cells. One of these genes it the arsenicreductase, arsC (gene ID: 2889). The aim of this study was to characterize thestructure of the ArsC proteinin order to determine the function of the gene. The prediction of the structure was conducted by usingthe online server I‐TASSER, which proposed a 3D model for the protein and predicted its role as anarsenate reductase. For verification of its function numerous laboratory techniques were used. The genewas isolated by PCR amplification using custom primers designed just for this project. The next step wasthe cloning of the gene into the pGEMT‐Easy vector and transformation of a mutant arsC‐ strain of E. coliOP50 with this vector containing the insert. After the transformation, the transformed colonies wereselected based on blue‐white screening and exposed to LB medium containing 50 mM sodium arsenate(As5+). The results obtained so far were not concluding, as no bacterial growth was observed in presenceof arsenate.

Place, publisher, year, edition, pages
2018.
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:his:diva-16376OAI: oai:DiVA.org:his-16376DiVA, id: diva2:1261993
Available from: 2018-11-09 Created: 2018-11-09 Last updated: 2018-11-09Bibliographically approved

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