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Rolling circle transcription on smallest size double stranded DNA minicircles
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
2010 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

The RNA polymerase T7 is utilized as a component of motor complexes in DNA nanotechnology due to its high promotor specificity, the lack of external transcription factors and its very high processivity, but there is no experience of its application on small double stranded DNA circles. Circular templates from 210 to 126 bp in circumference sharing a common promotor termination motif were synthesized and transcription was monitored at end point on gel and in real time with a 2’ O methyl RNA molecular beacon. The RNAP T7 was found to be able to utilize circular dsDNA templates down to 126 bp with moderate impact on transcription rate for saturated systems and rolling circle transcription products were evident with denaturizing PAGE gel electrophoresis for templates down to 167 bp.

Place, publisher, year, edition, pages
2010. , 63 p.
Series
UPTEC X, 10 026
Keyword [en]
RNA polymerase, RNAP T7, DNA nanotechnology, molecular motor, real time monitoring of transcription, 2’ O methyl RNA, molecular beacon, minicircle
National Category
Other Industrial Biotechnology
Identifiers
URN: urn:nbn:se:uu:diva-134082OAI: oai:DiVA.org:uu-134082DiVA: diva2:371559
Uppsok
Technology
Supervisors
Examiners
Available from: 2011-03-09 Created: 2010-11-21 Last updated: 2011-03-09Bibliographically approved

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Kristoffersson, Anders
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Biology Education Centre
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