Autonomously replicating plasmids carrying theAMA1 region inPenicillium chrysogenum |
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Authors: | Francisco Fierro Katarina Kosalková Santiago Gutiérrez Juan F. Martín |
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Affiliation: | (1) Area of Microbiology, Faculty of Biology, University of León, E-24071 León, Spain;(2) Institute of Biotechnology, INBIOTEC, E-24071 León, Spain |
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Abstract: | Plasmid vectors containing theAMA1 sequence transformed with high efficiency and replicated autonomously inPenicillium chrysogenum. The efficiency of transformation ofP. chrysogenum was related to the length of theAMA1 fragment used for constructing the different autonomously replicating plasmids. One of the two palindromic inverted repeats ofAMA1 (the 2.2-kbSalI-HindIII fragment) is sufficient to confer autonomous replication and a high transformation efficiency. Deletion of the 0.6-kb central fragment located between the inverted repeats did not affect either the ability of the plasmids to replicate autonomously or the efficiency of transformation, but did alter the mitotic stability and the plasmid copy number. Deletion of any fragment of the 2.2-kb repeat caused the loss of the ability to replicate autonomously and reduced the transformation efficiency. Most of the transformants retained the original plasmid configuration, as multimers and without reorganization, after several rounds of autonomous replication. TheAMA1 region works as an origin of replication inP. chrysogenum andA. nidulans but not apparently inAcremonium chrysogenum. |
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Keywords: | Filamentous fungi Origins of replication Stability Plasmid multimers |
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