Positive and negative selection using the tetA-sacB cassette: recombineering and P1 transduction in Escherichia coli.
Positive and negative selection using the tetA-sacB cassette: recombineering and P1 transduction in Escherichia coli.
复制标题
DOI：
10.1093/nar/gkt1075
复制
发表时间：
2013-12
影响因子：
14.9
通讯作者：
Court DL
中科院分区：
文献类型：
--
作者：
Li XT;Thomason LC;Sawitzke JA;Costantino N;Court DL
选择和反选择的两步过程是使用同源重组方法实现细菌基因组的遗传修饰和工程化的标准方式。tetA和sacB基因包含在一个DNA盒中，并赋予一种新的双重反选择系统。tetA的表达赋予细菌对四环素（TcR）的抗性，并且还引起对亲脂螯合剂镰刀菌酸的敏感性; sacB引起对蔗糖的敏感性。这两个基因通过TcR选择作为联合DNA盒引入大肠杆菌中。已经开发了一种含有镰刀菌酸和蔗糖的培养基，其中tetA-sacB的共表达作为反选择剂比单独的任一基因更敏感。结合重组工程和P1转导的同源重组方法，这种强大的系统已被用于选择细菌基因组中不能被其他反选择系统直接检测到的变化。
The two-step process of selection and counter-selection is a standard way to enable genetic modification and engineering of bacterial genomes using homologous recombination methods. The tetA and sacB genes are contained in a DNA cassette and confer a novel dual counter-selection system. Expression of tetA confers bacterial resistance to tetracycline (TcR) and also causes sensitivity to the lipophillic chelator fusaric acid; sacB causes sensitivity to sucrose. These two genes are introduced as a joint DNA cassette into Escherichia coli by selection for TcR. A medium containing both fusaric acid and sucrose has been developed, in which, coexpression of tetA-sacB is orders of magnitude more sensitive as a counter-selection agent than either gene alone. In conjunction with the homologous recombination methods of recombineering and P1 transduction, this powerful system has been used to select changes in the bacterial genome that cannot be directly detected by other counter-selection systems.
登录
查看更多内容
影响因子：
3.2
作者：
GAY, P;LECOQ, D;HOCH, JA
通讯作者：
HOCH, JA
影响因子：
3.6
作者：
BLOMFIELD, IC;VAUGHN, V;EISENSTEIN, BI
通讯作者：
EISENSTEIN, BI
影响因子：
5.6
作者：
Sawitzke JA;Costantino N;Li XT;Thomason LC;Bubunenko M;Court C;Court DL
通讯作者：
Court DL
DOI：
10.1073/pnas.121164898
发表时间：
2001-06-05
影响因子：
11.1
作者：
Ellis, HM;Yu, DG;Court, DL
通讯作者：
Court, DL
影响因子：
14.9
作者：
DeVito JA
通讯作者：
DeVito JA