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Rescue and characte...
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Murén, EvaUppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi
(author)
Rescue and characterization of episomally replicating DNA from the moss Physcomitrella
- Article/chapterEnglish2009
Publisher, publication year, extent ...
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2009-11-17
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Proceedings of the National Academy of Sciences,2009
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printrdacarrier
Numbers
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LIBRIS-ID:oai:DiVA.org:uu-109191
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-109191URI
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https://doi.org/10.1073/pnas.0908037106DOI
Supplementary language notes
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Language:English
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Summary in:English
Part of subdatabase
Classification
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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The moss Physcomitrella is unique among plants in that it permits efficient gene targeting by homologous recombination. Furthermore, transformed DNA can replicate episomally in Physcomitrella. Here we show that episomally replicating DNA can berescued back into E. coli, and use such rescue to study the fate of the transformed DNA. Significantly, plasmids rescued from moss transformed with circular DNA are identical to the original plasmid, whereas plasmids rescued from moss transformed with linearized DNA frequently have deletions created by direct repeat recombination.These events are highly predictable in that they target the longest direct repeat on the plasmid, if this repeat is at least 12 bp. Episomal transformants obtained with linearized DNA show a more than 1000-fold amplification of the DNA whereas transformants obtained with circular DNA have much lower copy numbers. Most episomal transformants quickly lose the plasmid in the absence of selection, but a semi-stable type of transformant that loses the plasmid at a much lower frequency was also observed. The consistent rescue of the original plasmid, or of predictable derivatives thereof, suggests that molecular genetics methods which rely on shuttle plasmids are feasible in Physcomitrella
Subject headings and genre
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Physcomitrella patens
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shuttle plasmid
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plasmid rescue
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recombination
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MEDICINE
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MEDICIN
Added entries (persons, corporate bodies, meetings, titles ...)
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Nilsson, AndersUppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi(Swepub:uu)annil103
(author)
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Ulfstedt, MikaelUppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi
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Johansson, MonikaUppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi(Swepub:uu)monjo282
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Ronne, HansUppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi(Swepub:uu)haron677
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Uppsala universitetInstitutionen för medicinsk biokemi och mikrobiologi
(creator_code:org_t)
Related titles
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In:Proceedings of the National Academy of Sciences of the United States of America: Proceedings of the National Academy of Sciences106:46, s. 19444-194490027-84241091-6490
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