. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ligund binding sites, the high level of expression in olfactory neurons, and the large size of the gene superfamily suggests that they may be olfactory receptors. In this study, we cloned members of the olfactory receptor genes from a permanently aquatic amphibian, the mudpuppy (Necturus nuiculnsus). and conducted a phylogenetic analysis of olfactory receptor genes from major vertebrate groups. Eighty previously published partial and full length olfactory receptor gene sequences from rat. mouse, human, dog. and cat- fish wer
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. ligund binding sites, the high level of expression in olfactory neurons, and the large size of the gene superfamily suggests that they may be olfactory receptors. In this study, we cloned members of the olfactory receptor genes from a permanently aquatic amphibian, the mudpuppy (Necturus nuiculnsus). and conducted a phylogenetic analysis of olfactory receptor genes from major vertebrate groups. Eighty previously published partial and full length olfactory receptor gene sequences from rat. mouse, human, dog. and cat- fish were aligned using the PileUp program of the GCG Se- quence Analysis Software Package (Genetics Computer Group. Inc). From two highly conserved regions in transmembrane do- mains 3 and 7, a pair of degenerate primers were designed: Tm3 5'-TGGCITAYGAYMGITAYGTKGC-3' 1ml .V-TADATRAAIGGRTTIARCAT-3' Total RNA was extracted with an Ultraspec RNA Isolation System (Biotex Inc.) from freshly dissected mudpuppy nasal tis- sue; it was treated with RQ1 RNase-free DNase (Promega), and converted to first strand cDNA with Superscript 11 Reverse Tran- scriptase (Gibco BRL). PCR was performed with an Ericomp thermocycler: the following parameters were used: 94°C for 2 niin. meandrtterence Dog_2 Chck_1 Figure 1. A phylogenetic tree derived from a neighbor-joining anal- ysis of olfactory receptor protein sequences. Bootstrap values greater than 80% are indicated. The accession numbers for the sequences are listed in the table below. The shaded region includes all fish sequences in the dataset as well as tH'o frog and one mudpuppy sequences that consistently cluster together. AND MOLECULAR BIOLOGY Title Genbank accession Title 249 Genbank accession Dog familiari.\ Rat Rtittu,\ itni'vt'xiiin Dog_J U53681 Rat_l M64376 Dog_2 U53679 Rat_2 M64377 Dog 3 U53680 Rat__3 M64378 Dog_4 U53682 Rat_4 M643S 1 Ra(_? M64386 Cattish Ictaltirus I'liiictalii.'i Rat 6 M64387 Catfish_l Catfish_2 Catfish_3 L092
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
