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. 2001 Feb 1;15(3):294–303. doi: 10.1101/gad.865401

Figure 2.

Figure 2

Additional mRNAs containing the SL sequence. (A) Amplification of multiple mRNAs by RT–PCR with SL and oligo(dT)-based primers. (Lane 1) Size markers [λ DNA, HindIII and EcoRI digest; in kilobases, from top to bottom, 21, 5.1/5.0, 4.2, 3.5, 2.0/1.9, 1.6, and (not visible) 1.4, 0.95, 0.83, and 0.5]. (Lanes 2,3) Body-wall muscle RNA template; (lanes 4,5) heart RNA template; (lane 6) no RNA. Reverse transcriptase was omitted in lanes 3 and 5. (B) 5′-untranslated sequences of three apparently complete mRNAs (GenBank AF237689–-AF237691) recovered by cloning DNA from the 1.0 and 0.8-kb bands in A, lane 2. The SL sequence is bolded; not shown is the BamHI site engineered at the 5′-end of the SL primer. The overlined ATG codons initiate ORFs of the lengths indicated, each terminated by a TAA stop codon and followed by an apparently complete 3′-untranslated sequence. In-frame stop codons within the 5′-untranslated sequences are underlined. The mRNA 1 ORF encodes a protein resembling HR-29 (Takagi et al. 1993), a myofibrillar protein from body wall muscle of the ascidian Halocynthia roretzi (46% identity over 207 aligned residues). The mRNA 2 ORF encodes a novel protein containing 17 PTDAVTL repeats resembling the mucin heptad [PTE(E/V)(P/T)TV] repeats of mammalian zonadhesins (Gao and Garbers 1998). The mRNA 3 ORF encodes a protein resembling vertebrate 27-kd heat-shock protein, hsp27 (Cooper and Uoshima 1994) (42% identity over 177 aligned residues).