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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 ?-glycosidase, and three carotenoid biosynthesis genes, CYP175A1, crtY, and crtI, from the genome of T. thermophilus HB27.

SUBMITTER: Wang L 

PROVIDER: S-EPMC4751836 | biostudies-other | 2015 Dec

REPOSITORIES: biostudies-other

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Markerless Gene Deletion with Cytosine Deaminase in Thermus thermophilus Strain HB27.

Wang Lei L   Hoffmann Jana J   Watzlawick Hildegard H   Altenbuchner Josef J  

Applied and environmental microbiology 20151211 4


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.  ...[more]

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