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. 2018 May 30;8:8383. doi: 10.1038/s41598-018-26560-9

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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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Cell-free synthesis of CLDN-5 with codon-converted mRNA. (a) Cell-free synthesized human CLDNs. Human CLDN-1 through CLDN-5 were synthesized on asolectin liposome using the wheat cell-free system with bilayer-dialysis method. After translation, proteoliposomes were purified by repeated centrifugation and resuspension in buffer. Upper panel: purified proteoliposome was subjected to SDS-PAGE and synthesized CLDNs were visualized by CBB staining. Arrowheads indicate translated CLDNs. Lower panel: gel electrophoresis of CLDN mRNAs. In vitro transcribed mRNA was applied to gel electrophoresis under non-denaturing conditions. (b) GC content of wild-type and codon-converted human CLDN5 mRNAs. The plot indicates local GC content, calculated by averaging 20 bases before and after each position. Figures in parentheses show total GC content. Black, wild-type CLDN5; red, codon-converted CLDN5.