Markerless Gene Deletion with Cytosine Deaminase in Thermus thermophilus Strain HB27

被引：5

作者：

Wang, Lei ^{[1
]}

Hoffmann, Jana ^{[1
]}

Watzlawick, Hildegard ^{[1
]}

Altenbuchner, Josef ^{[1
]}

机构：

[1] Univ Stuttgart, Inst Ind Genet, D-70174 Stuttgart, Germany

来源：

APPLIED AND ENVIRONMENTAL MICROBIOLOGY | 2016年 / 82卷 / 04期

关键词：

BETA-CAROTENE HYDROXYLASE; HOST-VECTOR SYSTEM; EXTREME THERMOPHILE; ESCHERICHIA-COLI; ALPHA-GALACTOSIDASE; DIRECTED EVOLUTION; SELECTION MARKER; CLONING VECTORS; SHUTTLE VECTORS; IN-VIVO;

D O I：

10.1128/AEM.03524-15

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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

引用

页码：1249 / 1255

页数：7

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