Open AccessAntiserum raised against gyrase A of Acholeplasma laidlawii reacts with phytoplasma proteins
Author(s) -
Koui Toru,
Natsuaki Tomohide,
Okuda Seiichi
Publication year - 2002
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2002.tb11004.x
Subject(s) - dna gyrase , antiserum , biology , antigenicity , fusion protein , escherichia coli , microbiology and biotechnology , gene , biochemistry , antigen , recombinant dna , genetics
Part of the gyrase A gene ( gyrA ) of Acholeplasma laidlawii was cloned and incorporated directly downstream from a 6×His tag segment of the pQE expression vector. The 23‐kDa fusion protein was expressed as a 6×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.