The species investigated were Pleuronectes platessa UGTIBl, P.platessa UGT,

Pleuronectes yokohamnae UGT IB2, Epinephelus coiodes UGT and Danio rerio UGT.

Five primers were designed which could hypothetically bind to this sequence. The

application of exclusion criteria (degeneracy- <100-fold, poor or no matches with fish

sequences resulting from BLASTn searches, %GC content <40%, potential to self-

dimerize < -20 kcal/mol) resulted in the selection of two primers, designated as UGTR3

and UGTR4, and chosen to be reverse primers (Table 5-1). An additional reverse primer

(UGT RS) was chosen due to its low degeneracy (4-fold) and its complementarity to the

highly conserved N-terminal domain downstream of the signature sequence. Five

additional primers (UGTF3-7) were also chosen based on these same criteria. Since

these primers were complementary to sequences upstream of the signature sequence, they

were selected to be forward primers (Table 5-1).

Table 5-1. 5' 3' Sequences of degenerate primers chosen.

ID Sequence Direction

UGT F3 GTGGTSCTGGT SCCYGAAASYAGY Forward

UGT F4 CTTACWGAYCCMTTCYTKCC STGYGGC Forward

UGT F5 AAC AT GGTYYWWATYGGRGGYAT CAAC TGT Forward

UGT F6 ATYGGRGGYATCAACTGTGCA Forward

UGT F7 GAGT TTGT SVAHGGC TCW GGA Forward

UGT R3 AAAC AGHGGRAACAT CAVC AT Reverse

UGT R4 YC CYT GS TCK SCAAAC AGHGG Reverse

UGT R5 GT GRTAC TGRAT C CAGTT CAG Reverse