20.500.12556/RUL-143717 A method for targeting a specified segment of DNA to a bacterial microorganelle Encapsulation of a selected DNA molecule in a cell has important implications for bionanotechnology. Non-viral proteins that can be used as nucleic acid containers include proteinaceous subcellular bacterial microcompartments (MCPs) that self-assemble into a selectively permeable protein shell containing an enzymatic core. Here, we adapted a propanediol utilization (Pdu) MCP into a synthetic protein cage to package a specified DNA segment in vivo, thereby enabling subsequent affinity purification. To this end, we engineered the LacI transcription repressor to be routed, together with target DNA, into the lumen of a Strep-tagged Pdu shell. Sequencing of extracted DNA from the affinity-isolated MCPs shows that our strategy results in packaging of a DNA segment carrying multiple LacI binding sites, but not the flanking regions. Furthermore, we used LacI to drive the encapsulation of a DNA segment containing operators for LacI and for a second transcription factor. DNA physical characterisation biochemical characterisation bakterijski mikrorazdelek Pdu Twin-Strep-tag represor LacI molekularna biologija biokemija true false true Angleški jezik Slovenski jezik Članek v reviji 2023-01-10 14:28:11 2023-01-10 14:28:12 2023-01-26 08:55:01 0000-00-00 00:00:00 2022 0 0 12 str. no. 19, art. e113 Vol. 50 2022 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 0305-1048 10.1093/nar/gkac714 120251907 gkac714.pdf gkac714.pdf 1 3A1FF3C37A5396C171777F93FBA2465D f03f526b11d86cb6ab8817eba9ba138f4054b5381d11c6edf7753651b1bbd0d4 e08c0d1f-924a-11ed-9c94-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=165703 https://academic.oup.com/nar/article/50/19/e113/6677558 0 b17500a1-90ea-11ed-9c94-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=165614 Biotehniška fakulteta 0 0 0