Markerless Gene Deletion with Cytosine Deaminase in Thermus thermophilus Strain HB27 | Zendy

Lei Wang | Zendy; Jana Hoffmann | Zendy; Hildegard Watzlawick | Zendy; Josef Altenbuchner | Zendy

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Markerless Gene Deletion with Cytosine Deaminase in Thermus thermophilus Strain HB27

Author(s) -

Lei Wang,

Jana Hoffmann,

Hildegard Watzlawick,

Josef Altenbuchner

Publication year - 2015

Publication title -

applied and environmental microbiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.552

H-Index - 324

eISSN - 1070-6291

pISSN - 0099-2240

DOI - 10.1128/aem.03524-15

Subject(s) - thermus thermophilus , biology , gene , plasmid , genetics , microbiology and biotechnology , escherichia coli

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.

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