Table I. Amassin peptide sequences obtained, and corresponding degenerate oligonucleotide primers used to clone cDNA.
| Peptide sequences (NH2 to COOH termini) | Synthetic oligonucleotides (5′ to 3′) |
|---|---|
| Trypsin | |
| DIIHVSEPFTVR (229–240) | CAYGTIWSIGARCCNTTYAC GTRAANGGYTCISWIACRTG |
| IGAWFRDPLQDYIK (246–259) | GGIGCNTGGTTYMGNGAYCC GGRTCNCKRAACCANGCICC |
| LDPETLDVIETWVAP (380–394) | GAYCCNGARACIYTNGAYGT ACRTCNARIGTYTCNGGRTC |
| LYGWDNGHQVVYDLTFDPPAR (460–480) | TGGGAYAAYGGNCAYCARGT ACYTGRTGNCCRTTRTCCCA |
| SQLLPTDTNIPLNLQ (481–495) | GAYACIAAYATHCCNYTIAA TTIARNGGDATRTTIGTRTC |
| Cyanogen bromide | |
| RTTSGVV (40–46) | |
| MTLEGVLDGTVVVTVAELELVKEELKEM (117–144) |
NH2-terminal peptide sequences obtained following trypsin or cyanogen bromide digestion. Peptide positions within the complete amassin protein sequence are noted in parentheses. Underlined amino acids represent the region to which the degenerate oligonucleotide primers were directed. Forward (top) and reverse (bottom) direction oligonucleotides were synthesized. Degenerate positions are denoted as follows: Y = C or T, N = A or C or G or T, R = A or G, I = inosine, M = A or C, K = G or T, W = A or T, S = C or G, H = A or C or T, D = A or G or T.