To establish SLAC1 complementation lines a 4.4 kb fragment including 1.63 kb of the 5′-UTR, the genomic SLAC1 gene region and 0.9 kb of the 3′-UTR (Negi et al., 2008) was amplified using the PfuX7 polymerase (Norholm, 2010).
Together with Box et al.'s [ 27] non-transgenic quantitative complementation assays, our transgenic complementations unequivocally establish the role of ELF3 in thermomorphogenesis signaling.
To establish a Cre-based complementation system ("split-Cre"), which allows recombination of LoxP-flanked DNA sequences depending on the simultaneous activity of two promoters (Fig. 1A), the coding sequence of Cre recombinase was cut into two complementation-competent fragments [15], [16] coding for amino acids residues 19 59 and 60 343.
In cell-based assays, we established that the complementation imaging reporter could detect CXCL12 binding to CXCR4 and quantify specific inhibition of ligand-receptor interaction.
Allelic combinations were established previously by complementation testing as described in [ 19] with the following exceptions.
Sequencing of dm3 mutants and transgenic complementation established RGC2B as the gene conferring Dm3 (Shen et al. 2002).
Using a lentiviral expression system, Calvo and colleagues [ 6] performed complementation experiments to establish pathogenicity of a mutation in NDUFB3 (NADH dehydrogenase (ubiquinone) 1 beta subcomplex, 3) carried by a patient with a complex I deficiency.
Further complementation studies are essential to establish the function of various isoforms of WRI in nonseed tissues.
To examine rigorously the ability of Nanog orthologs to establish naive pluripotency, we undertook a genetic complementation experiment (supplementary material Fig. S1A).
After tagging 114 essential genes using RMCE as described above, we performed complementation tests with null mutations or deletions to establish the fraction of tagged genes that failed to complement a severe loss of function or null allele.
