cDNA cloning and expression of Bauhinia purpurea lectin.
Bauhinia purpurea lectin (BPA) was purified from seeds of B. purpurea alba. The purified lectin was digested with an endoproteinase, Asp-N, or trypsin and then the amino acid sequences of the resultant fragments were analyzed. Furthermore, a cDNA library for BPA was constructed using RNA isolated from germinated Bauhinia purpurea seeds. By gene cloning, the nucleotide sequence of BPA cDNA and its deduced amino acid sequence were analyzed. The cloned BPA cDNA comprised 1,152 nucleotides and the open reading frame of the cDNA encodes a polypeptide of 290 amino acids including a signal peptide composed of 28 amino acids. BPA expressed in Escherichia coli showed a relative molecular mass of 29 kDa on sodium dodecyl sulfate-polyacrylamide gel. On comparison of its sequence with those of other leguminous seed lectins, BPA showed high homology to the others.
Kusui
K
K
Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical Sciences, University of Tokyo.
Yamamoto
K
K
Konami
Y
Y
Osawa
T
T
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004251:DNA Transposable Elements; D004591:Electrophoresis, Polyacrylamide Gel; D005690:Galactose / Q000737:chemistry; D015870:Gene Expression; D015723:Gene Library; D006868:Hydrolysis; D037102:Lectins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D037121:Plant Lectins; D010944:Plants / Q000737:chemistry*; D010957:Plasmids; D012710:Serum Albumin, Bovine / Q000737:chemistry
D004251:DNA Transposable Elements; D037102:Lectins; D037121:Plant Lectins; D012710:Serum Albumin, Bovine; D004247:DNA; D005690:Galactose