A Bradyrhizobium japonicum expression library based on phagemid vector pG3DSS was used to identify and isolate a large number of clones encoding secreted proteins (Rosander et al. 2003, MPMI 16, 727-737). N-terminal parts of B. japonicum proteins fused to the artificial E-tag peptide were detected in a phage display system by a highly specific monoclonal antibody. Following PCR amplification, some selected gene fusions were used to construct B. japonicum gene disruption mutants which were subsequently inoculated on soybean seedlings to test their symbiotic properties. While some of these mutants did not exhibit altered phenotypes, other mutants were severely affected in their symbiotic properties.
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