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. 2005 May;73(5):3128–3136. doi: 10.1128/IAI.73.5.3128-3136.2005

FIG. 1.

FIG. 1.

Flow chart of steps describing the identification of the initial 6.1 clone through the DNA sequencing of the 26-kb Bva-Brp fosmid clone. (1) Bartonella strain na19103nm genomic lambda library screened with antiserum from a mouse infected with Bartonella strains isolated from Peromyscus and Spermophilus reveals several positive plaques. A positive clone, 6.1, was selected for further characterization. (2) Western blot of the 58-kDa recombinant protein expressed by clone 6.1 reacted with the antiserum used in the library screening in step 1. Lane A, Escherichia coli with expression vector with no insert; lane B, E. coli with the 6.1 cloned insert. (3) The 1.5-kb insert of 6.1 is sequenced. Repeat regions are recognized. (4) PCR primers are synthesized from the 6.1 termini and used for amplification from Bartonella species genomic DNA. A similar-sized amplicon is observed from B. vinsonii subsp. arupensis (lane d). Lanes: a, B. quintana; b, B. henselae; c, B. vinsonii subsp. berkhoffii; d, B. vinsonii subsp. arupensis; e, B. elizabethae; f, B. bacilliformis; g, strain na19103 nm; h, clone 6.1. (5) B. vinsonii subsp. arupensis fosmid library is hybridized with the clone 6.1 probe to identify large inserts harboring analogous region. Positive clones are selected for analysis. (6) A B. vinsonii subsp. arupensis fosmid clone containing a 26-kb insert, Bva-Brp, was sequenced and analyzed.