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)