104



Scores

Seqik Name Len(aa) SeqB Name Len(aa) Score


1 livUGTp 627 2 I35RCp 133 60


livUGTp GMLRARVPGLFFSHSSSFFFSSKICFL-T- LETFKLKMPRLLAALCLQIYLCSFLGPVEG 58
I35R Cp

livUGTp GKVLVMPVDGSHWLSMKI LVEELSRRGHEMVVLVPETSVLIHGSDAYVARS FKVPYTKAE 118
I35R Cp

livUGTp LDESMNKLKEGITKAPRI SDLLENIIGLLS FTNMQVKGCEALLYNEPLMQNLREEHFDLM 178
I35R Cp

livUGTp LTDPFLPCGPI IAEAFSLPAVYFLRGLPCGLDLEAAQCPSPPSYVPRFFTGNTVTS 238
I35R Cp

livUGTp RVKNVLMTGFESILCKI FFSSFDELTSRYLKKDVTFRDVLGHAAIWLYRYDFTFEYPRP 298
I35R Cp

livUGTp MPNAVRIGGINCAKKNPLPADLEEFVDGSGDHGFIVFTLGSFVSELPEFKAREFFEAFRQ 358
I35R Cp -- -- -- -- -- -- -- -- -- -- -- -- -- -PANTTW ILWS KTHTEGH -- -HTVPVLNSVKI 2 8

livUGTp IPQRVLWRYTGVIPKDIPENVKVMKWLPQNDLL AHPKAKVFTGAGYICVP 417
I35R Cp -----I-KIRGNI- -MVHNNNFLFSFLSSPC-KAHPKAKVFITHGGTHGIYEGICGP 79

 t---------- UDPGA binding site ---
livUGTp VMIPLFGDQVDNVLRMVLRGVAESLTMFDLTSEQLLGALRKLNREITSLH 477
I35R Cp VMIPLFGDQVDNVLRMVLREVAESLTMFDLTSEQLLGALRKVNE-K--------- 128

livUGTp DRPIEPLDLAVFWTEFVMRHGSAEHLRPAAHHLNWVQYHSLVALVSTFIK 537
I35R Cp
 4-- TM region--~
livUGTp TCALCFRKCFRRAQKSKKE-NGQ-MIRNGFGAVFN-RRWFIVSCNKLSHV 592
I35R Cp

livUGTp QFNIQGCQQLWFSHCSYGCYDVTKKKK 619
I35R Cp
Figure 5-19. Alignment of predicted protein sequences from cloned catfish UGTs.
 Regions of interest and the starting and ending residue of the mature product
 are highlighted.


Cloning of entire UGT gene

 The liver UGT gene was successfully amplified and cloned, using sets of primers


that were complementary to both the untranslated regions (UTRs) upstream and


downstream of the gene, as well as primers that annealed to the exact start and end of the


translated portion of the gene (Figure 5-20). The same PCR conditions were used in an