title: Markerless Gene Deletion with Cytosine Deaminase in Thermus thermophilus Strain HB27.
abstract: We developed a counterselectable deletion system for Thermus thermophilus HB27 based on cytosine deaminase (encoded by codA) from Thermaerobacter marianensis DSM 12885 and the sensitivity of T. thermophilus HB27 to the antimetabolite 5-fluorocytosine (5-FC). The deletion vector comprises the pUC18 origin of replication, a thermostable kanamycin resistance marker functional in T. thermophilus HB27, and codA under the control of a constitutive putative trehalose promoter from T. thermophilus HB27. The functionality of the system was demonstrated by deletion of the bglT gene, encoding a β-glycosidase, and three carotenoid biosynthesis genes, CYP175A1, crtY, and crtI, from the genome of T. thermophilus HB27.
authors.affiliation: Institut für Industrielle Genetik, Universität Stuttgart, Stuttgart, Germany.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Institut für Industrielle Genetik, Universität Stuttgart, Stuttgart, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Hoffmann
authors.affiliation: Institut für Industrielle Genetik, Universität Stuttgart, Stuttgart, Germany.
authors.forename: Hildegard
authors.initials: H
authors.lastname: Watzlawick
authors.affiliation: Institut für Industrielle Genetik, Universität Stuttgart, Stuttgart, Germany josef.altenbuchner@iig.uni-stuttgart.de.
authors.forename: Josef
authors.initials: J
authors.lastname: Altenbuchner
mesh_terms: D000068016:Eubacteriales / Q000201:enzymology / Q000235:genetics; D043525:Cytosine Deaminase / Q000235:genetics / Q000378:metabolism*; D005437:Flucytosine / Q000633:toxicity*; D017353:Gene Deletion; D055786:Gene Knockout Techniques / Q000379:methods*; D005827:Genetics, Microbial / Q000379:methods*; D012641:Selection, Genetic; D016963:Thermus thermophilus / Q000235:genetics*; D001617:beta-Glucosidase / Q000235:genetics
chemical_list: D005437:Flucytosine; D001617:beta-Glucosidase; D043525:Cytosine Deaminase