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J Gen Virol. 2001 Jan;82(Pt 1):9-15.

Rapid identification of a tobacco mosaic virus epitope by using a coat protein gene-fragment-pVIII fusion library.

Author information

1
Institut für Biologie I (Botanik/Molekularbiologie), RWTH Aachen, Worringerweg 1, D-52074 Aachen, Germany.

Abstract

This study describes the identification of the epitope recognized by the tobacco mosaic virus (TMV) coat protein (CP)-specific monoclonal antibody 29 (MAb29) by displaying a CP gene-fragment library on pVIII of filamentous phage M13. More than 80% of the clones isolated after one round of panning bound specifically to MAb29. DNA sequencing of ten randomly chosen MAb29-specific clones and subsequent sequence comparison revealed a common seven amino acid epitope (ELIRGTG) representing amino acids 131-137 of the TMV CP. The reactivity of MAb29 in competition ELISA towards glutathione S:-transferase fused to this epitope was stronger than that towards full-length wild-type TMV CP, confirming the epitope sequence determined by gene-fragment phage display. This demonstrated that gene-fragment libraries displayed on the phage surface as fusion proteins with the filamentous bacteriophage gene VIII are useful tools for rapid identification of linear epitopes recognized by MAbs.

PMID:
11125152
DOI:
10.1099/0022-1317-82-1-9
[Indexed for MEDLINE]

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