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J Biosci ; 33(1): 91-101, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18376074

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).


Subject(s)
Gene Expression , Nyctaginaceae/genetics , Plant Proteins/genetics , Ribosome Inactivating Proteins/genetics , Amino Acid Sequence , Antiviral Agents/chemistry , Antiviral Agents/isolation & purification , Base Sequence , Cloning, Molecular , DNA, Complementary/genetics , Escherichia coli/genetics , Genes, Plant , Glycoside Hydrolases/analysis , Glycoside Hydrolases/genetics , Glycoside Hydrolases/metabolism , Molecular Sequence Data , Nyctaginaceae/anatomy & histology , Nyctaginaceae/chemistry , Open Reading Frames , Phylogeny , Plant Leaves/chemistry , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Leaves/metabolism , Plant Proteins/chemistry , 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/isolation & purification , Ribosome Inactivating Proteins/metabolism , Tobacco Mosaic Virus/physiology
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