Molecular Cloning of Nuclear Receptors

KIF17-T^ΔNR, KIF17-T^NR peptide, ERR1 and ERR3 were amplified by PCR from human Caco2 cells and cloned into Gateway^TM expression vectors (Invitrogen). Primers: KIF17-T^ΔNR: forward; 5’-GGG GAC AAG TTT GTA CAA AAA AGC AGG CTC CGA GCA GGT GCA GCC CCT GAT TC and reverse; GGG GAC CAC TTT GTA CAA GAA AGC TGG GTC TCA CAG AGG CTC ACT GCC AAA GTT-3’. KIF17-T^NR peptide: forward; 5’-GGG GAC AAG TTT GTA CAA AAA AGC AGG CTC CCA GGA GCG TGA CTC CAT GC and reverse; GGG GAC CAC TTT GTA CAA GAA AGC TGG GTC CAG GAG CTG CTG CAA GAG C-3’. ERR3: forward; 5’- GGG GAC AAG TTT GTA CAA AAA GCA GGC TCC ATG TCA AAC AAA GAT CGA C and reverse; GGG GAC CAC TTT GTA CAA GAA AGC TGG GTC GAC CTT GGC CTC CAA CAT T-3’. The KIF17-Tail and KIF1A-Tail constructs were described previously [40 (link)], [69 (link)]. ERR1: forward; 5’ - GTA CAA AAA AGC AGG CTT CTC CAG GGA GGT GGT GGG C and reverse; GTA CAA GAA AGC TGG GTC TCA GTC CAT CAT GGC CTC GAG. ERRE-Luc reporter was provided by JM Vanacker (University of Lyon, France). 3X-ERE-Luc was purchased from Addgene (Plasmid 11354). All constructs were verified by sequencing prior to use in experiments.