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. 2017 Oct 27;7:14246. doi: 10.1038/s41598-017-14475-w

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© The Author(s) 2017

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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Analysis of modified PGCs and sperm of recipient rooster. (a) CB PGCs were transfected with a mCherry expression plasmid consisting of mCherry under a chicken β-actin promoter and a puromycin resistance gene under a CAG promoter flanked by HS4 insulators. (b) Succesful selection and expression of mCherry was confirmed by fluorescence microscopy and (c) flow cytometry. (d) The DNA junction between the mCherry vector and adjacent chicken genomic sequence detected in the whole-genome sequence of parental PGC clone. The vector-derived attB sequence is in bold and the repetitive motifs in the chicken genomic DNA are underlined. (e) mCherry positivity in the spermiogenic epithelium of G₀ recipient rooster after orthotopic transplantation of mCherry-positive CB PGCs. (f) The mCherry reporter gene was detected by PCR in mCherry+ PGCs and semen samples of two recipient roosters.