Cloning and expression of antiviral/ribosome-inactivating protein from Bougainvillea xbuttiana.
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).
Choudhary
Nandlal
N
Division of Biochemistry, Indian Agricultural Research Institute, New Delhi 110012, India.
Kapoor
Harish C
HC
Lodha
Madan L
ML
D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000737:chemistry / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D006026:Glycoside Hydrolases / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D039281:Nyctaginaceae / Q000033:anatomy & histology / Q000737:chemistry / Q000235:genetics*; D016366:Open Reading Frames; D010802:Phylogeny; D018515:Plant Leaves / Q000737:chemistry / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D054788:Ribosome Inactivating Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014027:Tobacco Mosaic Virus / Q000502:physiology
D000998:Antiviral Agents; D018076:DNA, Complementary; D010940:Plant Proteins; D021382:Protein Sorting Signals; D011500:Protein Synthesis Inhibitors; D011993:Recombinant Fusion Proteins; D006026:Glycoside Hydrolases; D054788:Ribosome Inactivating Proteins