Qiack gel extraction kit

Plasmid and genomic DNA were isolated with Qiaprep Miniprep (Qiagen GmbH, Hilden, Germany) and Ultraclean Microbial DNA Isolation Kit (MoBio Laboratories, Carlsbad, CA, USA), respectively. When needed, DNA was also purified from agarose gels using a Qiack Gel extraction kit (Qiagen). DNA sequencing was performed by the dideoxy method at the Sequencing Unit of Centro de Investigación Médica Aplicada (CIMA, Universidad de Navarra, Spain), and primers were synthesized by Sigma-Aldrich Ltd. Searches for DNA and protein homologies were carried out using the Kyoto Encyclopedia of Genes and Genomes [[21 ]], EMBL-European Bioinformatics Institute server [[22 ]] and National Center for Biotechnology Information (NCBI) database [[23 ]].

Sequence data were obtained from Kyoto Encyclopedia of Genes and Genomes (KEGG²). Searches for DNA and protein homologies between Brucella species and other α-proteobacteria such as Ochrobactrum, Rhizobium, or Agrobacterium were carried out using KEGG, Basic Local Alignment Sequence Tool (BLAST³), and Clustal Omega⁴ from the European Molecular Biology Laboratory – European Bioinformatics Institute (EMBL-EBI⁵). New glycosyltransferase identification, using B. abortus 2308 was supported by Carbohydrate-Active enZymes database (CAZy⁶). Primers were designed using Primer 3 input⁷ and synthesized by Sigma–Aldrich. Plasmid DNA was extracted with Qiaprep spin Miniprep (Qiagen GmbH). When needed, DNA was purified from agarose gels using Qiack Gel extraction kit (Qiagen) and sequenced by the Servicio de Secuenciación of CIMA.

Genomic sequences were obtained from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database¹. Searches for DNA and protein homologies were carried out using the National Center for Biotechnology Information (NCBI²) and the European Molecular Biology Laboratory (EMBL) – European Bioinformatics Institute server³. Primers were synthesized by Sigma-Genosys (Haverhill, United Kingdom). DNA sequencing was performed by the “Servicio de Secuenciación del Centro de Investigación Médica Aplicada” (Pamplona, Spain). Restriction–modification enzymes were used under the conditions recommended by the manufacturer. Plasmid and chromosomal DNA were extracted with Qiaprep Spin Miniprep (Qiagen) and Ultraclean Microbial DNA Isolation Kits (Mo Bio Laboratories), respectively. When needed, DNA was purified from agarose gels using the Qiack Gel Extraction Kit (Qiagen).