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. 2024 Mar 23;14:38. doi: 10.1186/s13578-024-01206-8

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Design and validation of SL7207 synthetic expression cassette strains. (A) Schematic illustrating components and location of validation primers for incorporation of a synthetic shRNA expression cassette into the xylA locus of Salmonella strain SL7207. Two primer sets were used to test that the cassette was integrated (red dot internal primer set) and was in the correct orientation (green dot integration primer set). (B) The expected PCR products of 274 bp (integration primers) and 444 bp (internal primers) for both the positive control and the scrambled (SCR) and c-Myc shRNA strains were observed. (C) Percentage of KmR cells five days (∼ 80 divisions) following kanamycin withdrawal demonstrating the retention of chromosomal (SL7207/Chr-KmR) versus vector (SL7207/pME6-KmR) derived resistance (**** P < 0.0001; unpaired T-test)