Structure/cdd/cdd. shtml; Marchler-Bauer and Bryant 2004), Emboss software

(http://www.bioweb .pasteur.fr) and CLC Protein Workbench v. 1.5.2 (http://www.clcbio.

com).

 Results (part 2)

 The RACE studies enabled the successful amplification and cloning of several

partial DNA sequences with good similarity to known UGTs and unknown fish

sequences. In the case of the liver, partial overlapping sequences obtained via cloning and

amplification of PCR products resulted in the recovery of the whole gene (livUGTn,

Figure 5-7), including untranslated regions at both ends. For the intestine, only partial

sequences, corresponding to two distinct isoforms were obtained (I4_3R and I35R PCR)

by RACE. However, the use of GSPs UTRF1 and UTRR2 resulted in the amplification

and cloning of an identical gene from intestine (intUGTn), which was identical to the 14-

3R partial sequence. The I35RPCR sequence was only 39% homologous with the full

length UGTs obtained from liver and intestine. A comparison of the relative sizes of

these sequences with respect to the full-length liver and intestinal UGT genes is shown in

Figure 5-8. For both liver and intestine truncated forms of UGT were identified

(L25R_5A, I4_6A, I35RPCR). These lacked the coding sequence for the trans-

membrane segment at the 3'-end of the gene.

 The sequences for the amplified, partial-sequence cDNAs that were sequenced

directly or when cloned are given in Appendix A.

Nucleotide sequence analysis

 a. Full-length UGT from liver/intestine. The UGTn sequence was subjected to a

BLASTn search (Table 5-7), showing that the sequence exhibited a high degree of

similarity to known UGT sequences. However, DNA sequences are inherently noisy (due