Cloning and expression of cDNA coding for bouganin

Eur J Biochem. 2002 Mar;269(6):1772-9. doi: 10.1046/j.1432-1327.2002.02823.x.

Abstract

Bouganin is a ribosome-inactivating protein that recently was isolated from Bougainvillea spectabilis Willd. In this work, the cloning and expression of the cDNA encoding for bouganin is described. From the cDNA, the amino-acid sequence was deduced, which correlated with the primary sequence data obtained by amino-acid sequencing on the native protein. Bouganin is synthesized as a pro-peptide consisting of 305 amino acids, the first 26 of which act as a leader signal while the 29 C-terminal amino acids are cleaved during processing of the molecule. The mature protein consists of 250 amino acids. Using the cDNA sequence encoding the mature protein of 250 amino acids, a recombinant protein was expressed, purified and characterized. The recombinant molecule had similar activity in a cell-free protein synthesis assay and had comparable toxicity on living cells as compared to the isolated native bouganin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Northern
  • Blotting, Western
  • Cell Line
  • Cloning, Molecular
  • DNA, Complementary
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Magnoliopsida / chemistry*
  • Molecular Sequence Data
  • N-Glycosyl Hydrolases / chemistry
  • N-Glycosyl Hydrolases / genetics*
  • N-Glycosyl Hydrolases / isolation & purification
  • Open Reading Frames
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Ribosome Inactivating Proteins
  • Sequence Homology, Amino Acid

Substances

  • DNA, Complementary
  • Recombinant Proteins
  • N-Glycosyl Hydrolases
  • Ribosome Inactivating Proteins

Associated data

  • GENBANK/AF445416