Cloning of a lectin cDNA and seasonal changes in levels of the lectin and its mRNA in the inner bark of Robinia pseudoacacia.
A cDNA clone encoding a lectin was isolated by immunological screening of an expression library prepared from poly(A)+ RNA from the inner bark of Robinia pseudoacacia. The cDNA clone (RBL104) had an open reading frame of 858 bp that encoded a polypeptide with a predicted molecular weight of 31210. This molecular weight corresponded closely to that of a polypeptide immunoprecipitated from products of translation in vitro of the poly(A)+ RNA. Thus, RBL104 appeared to be a full-length cDNA. The N-terminal amino acid sequence of the purified lectin protein matched a portion of the predicted amino acid sequence. It appeared that the lectin was synthesized as a precursor that consisted of a putative signal peptide of 31 amino acids and a mature polypeptide of 255 amino acids. Southern blot analysis of the genomic DNA revealed that the lectin was encoded by a small multigene family. The lectin was mostly localized in the axial and ray parenchymal cells of the inner bark. A small amount of lectin was also found in the axial and ray parenchymal cells of the xylem. The lectin accumulated in the inner bark in September, remained at high levels during the winter and disappeared in May. The mRNA for the lectin was detected from August to the following March. The appearance and disappearance of the mRNA were observed prior to those of the lectin protein.
Yoshida
K
K
Forestry and Forest Products Research Institute, Ibaraki, Japan.
Baba
K
K
Yamamoto
N
N
Tazaki
K
K
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D007887:Fabaceae / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D037102:Lectins / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D037121:Plant Lectins; D010946:Plants, Medicinal; D011061:Poly A / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis*; D012621:Seasons; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D018076:DNA, Complementary; D037102:Lectins; D037121:Plant Lectins; D012333:RNA, Messenger; C047720:Robinia lectin; D011061:Poly A