Molecular cloning of the antibacterial protein of the giant African snail, Achatina fulica Férussac.
An expression cDNA library was constructed with poly(A)-rich RNA extracted from the collar of the giant African snail, Achatina fulica Férussac. A 1.9-kbp cDNA clone encoding a precursor of antibacterial glycoprotein of the snail, achacin, was isolated from the cDNA expression library. The cDNA sequence contains an open reading frame with 1593-nucleotide residues. The deduced amino acid sequence of this achacin precursor starts with a 29-residue leader peptide followed by a 502-residue mature peptide (56 kDa) with four possible N-glycosylation sites, Asn-Xaa-Ser or Asn-Xaa-Thr. The Northern-blot analysis proved that the achacin precursor was specifically expressed in the tissue of snail collar and processed to mature achacin. cDNA inserts encoding achacin precursor were subcloned into expression plasmids. Three kinds of expressed polypeptides were cross-reacted with rabbit antiserum raised against achacin. The largest polypeptide (M(r) 63,000) should be the achacin precursor.
Obara
K
K
Department of Chemistry, Faculty of Science and Technology, Sophia University, Tokyo, Japan.
Otsuka-Fuchino
H
H
Sattayasai
N
N
Nonomura
Y
Y
Tsuchiya
T
T
Tamiya
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D003062:Codon; D004247:DNA / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006031:Glycosylation; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009479:Neuropeptides / Q000737:chemistry / Q000235:genetics*; D010446:Peptide Fragments / Q000737:chemistry; D010957:Plasmids; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics; D012697:Serine Endopeptidases; D012908:Snails / Q000737:chemistry* / Q000235:genetics
D003062:Codon; D009479:Neuropeptides; D010446:Peptide Fragments; D011498:Protein Precursors; C077200:achacin; D004247:DNA; D012697:Serine Endopeptidases; C036813:lysyl endopeptidase