title: cDNA cloning and in vitro synthesis of the Dolichos biflorus seed lectin.
abstract: The Dolichos biflorus seed lectin contains two structurally related subunits. A cDNA library was constructed using RNA isolated from D. biflorus seeds actively synthesizing the seed lectin. The library was expressed in Escherichia coli using a lambda Charon 16 vector, and lectin-specific antiserum was used to isolate a seed lectin cDNA. Hybridization of the D. biflorus seed lectin cDNA to RNA isolated from seeds actively producing both lectin subunits identifies a single-size RNA of 1100 bases. An oligodeoxyribonucleotide probe, constructed from an amino acid sequence common to both lectin subunits, detects the same size RNA. Translation of seed mRNA in vitro and immunoprecipitation of translation products using a lectin-specific antiserum yields a single polypeptide of slightly higher molecular mass than the largest seed lectin subunit. This seed lectin precursor is indistinguishable from a polypeptide synthesized from mRNA hybrid selected by the seed lectin cDNA. These data support the existence of a single polypeptide precursor for both subunit types of the D. biflorus seed lectin and suggest that differences between the subunit types arise by posttranslational processing.
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D037102:Lectins / Q000096:biosynthesis / Q000235:genetics*; D037121:Plant Lectins; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D037102:Lectins; D037121:Plant Lectins; D012333:RNA, Messenger; C036554:dolichos biflorus agglutinin; D004247:DNA