WebMay 24, 2024 · We tested counter-selection efficiency in multiple Salmonella loci using the commonly used markers sacB or I-sceI (Fig. 1b). sacB codes for levan sucrase, which … WebNov 6, 2013 · When Tet/SacB counter-selection agar is used during a P1 transduction , it is modified by adding 5 ml of a sterile 1 M sodium citrate solution to the agar for a final concentration of 5 mM. This is the P1 Tet/SacB counter-selection agar. Creating the tetA-sacB counter-selection cassette.
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WebMar 20, 2024 · The selection stringency of kil was measured and compared with the most widely used counter-selection gene, sacB, at the lacI, ack, and dbpa loci using different E. coli strains. At all these loci of tested strains, the selection stringency of kil significantly exceeds that of sacB by 2- to 28-fold. When dsDNA fragments were employed for ... WebMay 30, 2024 · In this study, we combine the sacB-based negative selection system with a cre-lox antibiotic marker recycling method. This methodology allows allelic exchange … filey live
A Modified Janus Cassette (Sweet Janus) to Improve Allelic
WebMar 20, 2024 · Seamless modification of the Escherichia coli genome using positive selection / negative selection is widely used in metabolic engineering and functional genome analysis. Some excellent negative selection systems have been reported, of which tetA-sacB and inducible toxins system are prominent. To expand the existing negative … WebFeb 27, 2012 · sacB is perhaps the most widely used counterselectable marker. The sacB gene was isolated from Bacillus subtilis and encodes the for the enzyme levansucrase. Expression of sacB in most gram-positive bacteria is harmless, but is lethal when expressed in gram-negative bacteria in the presence of sucrose [ 2 ]. WebI have a problem getting my sacB counter-selection to work. sacB sequence (obtained from PYUB870) was cloned into the MCS of pGEM-T, transformed into E. coli and plated on 1) 5% sucrose LB agar ... groovy final