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. 2016 Apr 14;16:37. doi: 10.1186/s12896-016-0267-8

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© Mukai et al. 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 2 — Generation of GFP knockin strain for carA-1. a Genomic organization of wild-type (WT) and GFP knockin locus for DDB_G0273397/carA-1. b WT and knockin locus before and after the removal of BsR cassette was detected by PCR. The primer set fw1/rev1, both located outside the homology arms, detects WT and knockin locus (pre_Cre) as 2.0 and 4.5 kb, respectively. BsR-removal was detected as the decrease in the size of target locus from 4.5 kb (pre-Cre) to 2.7 kb (post-Cre). Primer combination of rev2 for GFP and fw1 confirms specific recombination at the 5′ arm by yielding a 1.0 kb fragment. c Expression of GFP-tagged cARA-1 protein of the knockin strain detected by western blotting. Lysate of cells over-expressing cARA-1-GFP from extrachromosomal plasmid were loaded as the detection control (1/10 volume, cARA-1-GFP O.E)