Antiserum raised against gyrase A of Acholeplasma laidlawii reacts with phytoplasma proteins.
Part of the gyrase A gene (gyrA) of Acholeplasma laidlawii was cloned and incorporated directly downstream from a 6 x His tag segment of the pQE expression vector. The 23-kDa fusion protein was expressed as a 6 x His-tagged protein in Escherichia coli. The fusion protein was purified and used as an antigen for rabbit immunization. Western immunoblot analysis revealed that the antiserum raised against the gyrase A fragment had a specific affinity for a 108-kDa protein of A. laidlawii and cross-reacted with a 107.5-kDa protein of Acholeplasma axanthum, a 107-kDa protein of Acholeplasma granularum, and 95-97-kDa proteins of several phytoplasma-infected plants. The antiserum could also detect phytoplasmas in infected plant sap. These results demonstrate that the gyrase A protein (GyrA) of A. laidlawii shares antigenicity with the GyrA of other Acholeplasma species and also with those of phytoplasmas including some from a few groups with unrelated 16S rRNAs.
Koui
Toru
T
Faculty of Agriculture, Utsunomiya University, Utsunomiya 321-8505, Japan.
Natsuaki
Tomohide
T
Okuda
Seiichi
S
D000127:Acholeplasma / Q000201:enzymology* / Q000276:immunology; D000128:Acholeplasma laidlawii / Q000201:enzymology* / Q000276:immunology; D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000276:immunology*; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular; D003429:Cross Reactions; D027081:DNA Gyrase / Q000235:genetics / Q000276:immunology*; D019684:Magnoliopsida / Q000382:microbiology; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D027081:DNA Gyrase