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Abstract: The widely used pSU8 family of cloning vectors is based on a p15A replicon and a chloramphenicol
acetyltransferase (cat) gene conferring chloramphenicol resistance.We frequently
observed an increase in the size of plasmids derived from these vectors. Analysis of the bigger
molecular species shows that they have an IS10 copy inserted at a specific site between
the promoter and the cat open reading frame. Promoter activity from both ends of IS10 has
been reported, suggesting that the insertion events could lead to higher CAT production.
Insertions were observed in certain constructions containing inserts that could lead to plasmid
instability. To test the possibility that IS10 insertions were selected as a response to
chloramphenicol selection, we have grown these constructs in the presence of different
amounts of antibiotic and we observed that insertions arise promptly under higher chloramphenicol
selective pressure. IS10 is present in many E. coli laboratory strains, so the possibility
of insertion in constructions involving cat-containing vectors should be taken into
account. Using lower chloramphenicol concentrations could solve this problem.
Fuente: PLoS One. 2015 Sep 16;10(9):e0138615
Editorial: Public Library of Science
Año de publicación: 2015
Nº de páginas: 9
Tipo de publicación: Artículo de Revista
CORAL GONZALEZ PRIETO
LETICIA AGUNDEZ CORTES
MATXALEN LLOSA BLAS