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. 2018 Jul 5;8:10223. doi: 10.1038/s41598-018-28217-z

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© The Author(s) 2018

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Detection of transposition activities of five common transposons in P. pastoris and liquid enrichment of transposon mutants. (A) A schematic diagram showing the two-component assay system. A helper plasmid integrated into the P. pastoris genome provides transposase under the control of an inducible promoter. A donor plasmid provides the Transposon Element (TE) lacking transposase but containing a HIS4⁺ selection marker. (B) YND plates after His⁺ transformant selection. Compared with pBRAmp-TIRsHis, pBRAmp-His lacks the corresponding TIRs which are necessary components for transposition to happen. (C) Integration assays of the five common transposons: Himar1, Mos1, Osmar14, SB and TcB in P. pastoris. “−” is the negative control which measures the transposition of a HIS4⁺ expression cassette from the pBRAmp-His into wild type genome. (D) Growth curves of GS115, GS115 + 1/10,000 GS115-GFP-HIS4, GS115 + 1/100 GS115-GFP-HIS4 and GS115-GFP-HIS4 cells.