Rational optimization of tolC as a powerful dual selectable marker for genome engineering

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Rational optimization of tolC as a powerful dual selectable marker for genome engineering

Selection has been invaluable for genetic manipulation, although counter-selection has historically exhibited limited robustness and convenience. TolC, an outer membrane pore involved in transmembrane transport in E. coli, has been implemented as a selectable/counter-selectable marker, but counter-selection escape frequency using colicin E1 precludes using tolC for inefficient genetic manipulat...

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Recombineering with tolC as a Selectable/Counter-selectable Marker: remodeling the rRNA Operons of Escherichia coli

This work describes the novel use of tolC as a selectable/counter-selectable marker for the facile modification of DNA in Escherichia coli. Expression of TolC (an outer membrane protein) confers relative resistance to toxic small molecules, while its absence renders the cell tolerant to colicin E1. These features, coupled with the lambdaredgam recombination system, allow for selection of tolC i...

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We have tested whether the thymidine kinase gene of herpes simplex virus 1 (HSV-1 tk) could function as a negative selectable marker in Leishmania major. Several drug resistance markers have been used for positive selection of transfected DNA in trypanosomatids, including neomycin phosphotransferase (NEO; refs. 1-4) and hygromycin phosphotransferase (HYG; refs. 5,6). Negative selectable markers...

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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 2014

ISSN: 1362-4962,0305-1048

DOI: 10.1093/nar/gkt1374