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

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Abstract

A full-length cDNA encoding ribosome-inactivating/antiviral protein (RIP/AVP) from the leaves of Bougainvillea xbuttiana 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 xbuttiana antiviral protein1). The ORF was cloned into an expression vector and expressed in E. coli as a fusion protein of ∼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).

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Abbreviations

AVP:

antiviral protein

BBAP1:

Bougainvillea xbuttiana antiviral protein1

BLAST:

basic local alignment search tool

ORF:

open reading frame

RIP:

ribosome-inactivating protein

RLM-RACE:

RNA ligase-mediated rapid amplified cDNA ends

RT-PCR:

reverse transcription polymerase chain reaction

SOD:

superoxide dismutase

TCS:

trichosanthin

TMV:

tobacco mosaic virus

UTR:

untranslated region

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Author notes

  1. Madan L. Lodha

    Present address: Emeritus Scientist, Division of Genetics, Indian Agricultural Research Institute, New Delhi, 110012, India

Authors and Affiliations

  1. Division of Biochemistry, Indian Agricultural Research Institute, New Delhi, 110012, India

    Nandlal Choudhary, Harish C. Kapoor & Madan L. Lodha

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  1. Nandlal Choudhary
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  2. Harish C. Kapoor
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  3. Madan L. Lodha
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Correspondence to Madan L. Lodha.

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Choudhary, N., Kapoor, H.C. & Lodha, M.L. Cloning and expression of antiviral/ribosome-inactivating protein from Bougainvillea xbuttiana . J Biosci 33, 91–101 (2008). https://doi.org/10.1007/s12038-008-0025-8

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  • DOI: https://doi.org/10.1007/s12038-008-0025-8

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