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.