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
收藏
页码：1249 / 1255
页数：7
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