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dc.contributor.authorBilyk, Oksana
dc.contributor.authorSekurova, Olga N
dc.contributor.authorZotchev, Sergey B
dc.contributor.authorLuzhetskyy, Andriy N
dc.date.accessioned2016-08-17T13:17:27Z
dc.date.available2016-08-17T13:17:27Z
dc.date.issued2016
dc.identifier.citationCloning and Heterologous Expression of the Grecocycline Biosynthetic Gene Cluster. 2016, 11 (7):e0158682 PLoS ONEen
dc.identifier.issn1932-6203
dc.identifier.pmid27410036
dc.identifier.doi10.1371/journal.pone.0158682
dc.identifier.urihttp://hdl.handle.net/10033/618497
dc.description.abstractTransformation-associated recombination (TAR) in yeast is a rapid and inexpensive method for cloning and assembly of large DNA fragments, which relies on natural homologous recombination. Two vectors, based on p15a and F-factor replicons that can be maintained in yeast, E. coli and streptomycetes have been constructed. These vectors have been successfully employed for assembly of the grecocycline biosynthetic gene cluster from Streptomyces sp. Acta 1362. Fragments of the cluster were obtained by PCR and transformed together with the "capture" vector into the yeast cells, yielding a construct carrying the entire gene cluster. The obtained construct was heterologously expressed in S. albus J1074, yielding several grecocycline congeners. Grecocyclines have unique structural moieties such as a dissacharide side chain, an additional amino sugar at the C-5 position and a thiol group. Enzymes from this pathway may be used for the derivatization of known active angucyclines in order to improve their desired biological properties.
dc.language.isoenen
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/281623en
dc.rightsopenAccessen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleCloning and Heterologous Expression of the Grecocycline Biosynthetic Gene Cluster.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1, 66123 Saarbrücken, Germany.en
dc.identifier.journalPloS oneen
refterms.dateFOA2018-06-12T17:47:09Z
html.description.abstractTransformation-associated recombination (TAR) in yeast is a rapid and inexpensive method for cloning and assembly of large DNA fragments, which relies on natural homologous recombination. Two vectors, based on p15a and F-factor replicons that can be maintained in yeast, E. coli and streptomycetes have been constructed. These vectors have been successfully employed for assembly of the grecocycline biosynthetic gene cluster from Streptomyces sp. Acta 1362. Fragments of the cluster were obtained by PCR and transformed together with the "capture" vector into the yeast cells, yielding a construct carrying the entire gene cluster. The obtained construct was heterologously expressed in S. albus J1074, yielding several grecocycline congeners. Grecocyclines have unique structural moieties such as a dissacharide side chain, an additional amino sugar at the C-5 position and a thiol group. Enzymes from this pathway may be used for the derivatization of known active angucyclines in order to improve their desired biological properties.


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