ABSTRACT
The movement protein (MP) of the tobacco mosaic virus (TMV) provides the intercellular transport of the viral RNA through plasmodesmata. The MP fulfills its function while interacting with host cell factors over the whole path of its intracellular movement from the subcellular site of its synthesis to the plasmodesmata of cellular walls. The MP conformation during its intracellular movement and fulfillment of the transport function still remains unknown. In this study, we describe the preparation of murine monoclonal antibodies (MAs) to TMV MP and mapping of the MP epitopes. Stable hybridoma lines that produce MAs to the partially denatured recombinant MP (MPr) were obtained. MAs were tested by immunoblotting and ELISA with the use of deletion variants of MPr. The epitopes of TMV MPr that recognize specific MAs were determined.
Subject(s)
Antibodies, Monoclonal , Epitope Mapping/methods , Recombinant Fusion Proteins/metabolism , Tobacco Mosaic Virus/metabolism , Viral Proteins/metabolism , Animals , Biological Transport , Cloning, Molecular , Endoplasmic Reticulum/metabolism , Mice , Plant Viral Movement Proteins , Recombinant Fusion Proteins/genetics , Nicotiana/virology , Viral Proteins/geneticsABSTRACT
Chimeric oligo(deoxyribo-ribo)nucleotides appeared to be a valuable tool to achieve the high selectivity of RNA cleavage as shown by RNA-ase H-mediated hydrolysis of TMV RNA directed by d(TGTGTATGCC), d(TGTGTAT), d(TGTGTAT)GCCAU and d(TGTGTAT)ppGCCAU.
