Complex and minimal media and culture growth conditions were as described previously (Dunn et al. 1996). Radiochemical assays to measure CO2 fixation from NaH14C03, and detection of BDCs on protein blots with streptavidin-horseradish peroxidase conjugate were performed as described previously (Dunn et al. 1996; M. Dunn, G. Araíza, T. Finan, submitted). Genome sequences putatively encoding BDCs in Mesorhizobium loti (Kaneko et al. 2000; http://www.kazusa.or.jp/rhizobase) were located by a keyword search followed by an open reading frame and BLASTP analyses of the sequences flanking the putative BDC. BDCs in the Sinorhizobium meliloti (http://sequence.toulouse.inra.fr/meliloti.html) genome were identified similarly, except that the identity of genes surrounding the BDCs were obtained using the Map function available at the site. Deduced genomic protein sequences of both genomes were also searched by BLASTP using various BDC sequences from other organisms as a query.
Was this article helpful?