Fig. 7. RDA clone 125-10 identifies the gene for a novel protein, tramdorin1. A, cDNAs that correspond to RDA clone 125-10. Five cDNA fragments are shown; they include the RDA fragment 125-10, two independent, nonidentical 5′ RACE rat cDNAs, a 3′ RACE rat cDNA, and a mouse EST cDNA. Of these, the 2.45 kb EST cDNA 1920302 appears to include the entire open reading frame, which is depicted in black. This cDNA appears to be nearly full length, based on the calculated size of tramdmRNA (Figs. 3, 7). The organism and tissue of origin of each cDNA fragment are listed. The open reading frame commences with an ATG initiation codon (in bold) that closely matches the Kozak consensus (Kozak, 1987); nucleotides that are an exact match areunderlined. 14dpc, 14 d postcoitum. B, Amino acid sequences encoded by mouse tramdorin EST cDNA 1920302 and a hypothetical rat tramdorin1 cDNA, derived from 5′ and 3′ RACE sequences, are shown. These sequences have been assigned GenBank accession numbers AF512429 and AF512430, respectively. The mouse cDNA encodes a 478 aa protein, whereas the corresponding rat protein is 481 aa. The amino acid sequence of mouse EST cDNA 1920302 was analyzed using the transmembrane domain prediction programs Memsat2 (McGuffin et al., 2000) and TMHMM (Sonnhammer et al., 1998). Eleven putative transmembrane domains are numbered. Transmembrane domains predicted by TMHMM are marked with capital Ts. For Memsat2, transmembrane domains are marked as follows: O, Outside transmembrane helix cap; X, central transmembrane helix segment; I, inside transmembrane helix cap. For both Memsat2 and TMHMM, predicted cytoplasmic domains are marked by +, whereas noncytoplasmic domains are marked by −. Five consensus glycosylation sites within the protein sequence are shown inbold and are boxed. C, Diagram depicting the predicted topology of tramdorin1 with 11 transmembrane domains, as predicted by Memsat2 and TMHMM. The three extracellular/lumenal glycosylation sites are marked with branched structures. However, the electrophoretic mobility of tramdorin suggests that it is not glycosylated extensively (Fig. 8).