Cloning and expression of antiviral/ribosome-inactivating protein from Bougainvillea xbuttiana

J Biosci. 2008 Mar;33(1):91-101. doi: 10.1007/s12038-008-0025-8.

Abstract

A full-length cDNA encoding ribosome-inactivating/antiviral protein (RIP/AVP)from the leaves of Bougainvillea x buttiana was isolated. The cDNA consisted of 1364 nucleotides with an open reading frame (ORF)of 960 nucleotides encoding a 35.49 kDa protein of 319 amino acids. The deduced amino acid sequence has a putative active domain conserved in RIPs/AVPs and shows a varying phylogenetic relationship to the RIPs from other plant species. The deduced protein has been designated BBAP1 (Bougainvillea x buttiana antiviral protein1). The ORF was cloned into an expression vector and expressed in E.coli as a fusion protein of approximately 78 kDa. The cleaved and purified recombinant BBAP1 exhibited ribosome-inhibiting rRNA N-glycosidase activity,and imparted a high level of resistance against the tobacco mosaic virus (TMV).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antiviral Agents / chemistry
  • Antiviral Agents / isolation & purification
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Escherichia coli / genetics
  • Gene Expression*
  • Genes, Plant
  • Glycoside Hydrolases / analysis
  • Glycoside Hydrolases / genetics
  • Glycoside Hydrolases / metabolism
  • Molecular Sequence Data
  • Nyctaginaceae / anatomy & histology
  • Nyctaginaceae / chemistry
  • Nyctaginaceae / genetics*
  • Open Reading Frames
  • Phylogeny
  • Plant Leaves / chemistry
  • Plant Leaves / genetics
  • Plant Leaves / growth & development
  • Plant Leaves / metabolism
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plant Proteins / isolation & purification
  • Plant Proteins / metabolism
  • Protein Sorting Signals
  • Protein Structure, Tertiary
  • Protein Synthesis Inhibitors / chemistry
  • Protein Synthesis Inhibitors / isolation & purification
  • Protein Synthesis Inhibitors / metabolism
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Ribosome Inactivating Proteins / chemistry
  • Ribosome Inactivating Proteins / genetics*
  • Ribosome Inactivating Proteins / isolation & purification
  • Ribosome Inactivating Proteins / metabolism
  • Tobacco Mosaic Virus / physiology

Substances

  • Antiviral Agents
  • DNA, Complementary
  • Plant Proteins
  • Protein Sorting Signals
  • Protein Synthesis Inhibitors
  • Recombinant Fusion Proteins
  • Glycoside Hydrolases
  • Ribosome Inactivating Proteins