melitensis 16 M . PCR amplification
of wbkE, manB O – Ag , manA O – Ag , manC O – Ag , wkdD, wbkF, wboA and wboB, wa** and manB core was conducted on representative strains of each of the Brucella species included in this study and their biovars with attention to the LPS characteristics (i.e. S versus R; and A dominant, M dominant, or A = M for the S-LPS). PD332991 Except for wboA and wboB in B. ovis, all genes were successfully amplified in the strains of all Brucella species and biovars tested. These results confirm the absence of the wbo selleck chemicals region in B. ovis [16,17]. They also suggest that conservation of wbk extends beyond those genes ( wbkA to wbkC ) examined in a previous work  and that wa** and manB core were are also conserved in the genus. Further analyses were then conducted to examine these possibilities. Gene polymorphism in wbk wbkE For all strains, the wbkE PCR-amplified product displayed the same Eco RV, Hinf I, Pst I and Pvu II RFLP patterns. Although B. melitensis 63/9 biovar 2 showed a different Sty I pattern, only one of eight additional B. melitensis biovar 2 strains tested showed this Sty I pattern (data not shown). manA O – Ag B. neotomae had a distinct manA O – Ag restriction pattern consisting of an additional Ava II site (Figures 2 and 3, Table 1). Moreover, in silico analysis showed a specific profile for B. ovis consisting
of a nucleotide substitution (GAA to GGA) at position 497 which modified the ManA C-terminal sequence Adenosine at amino acid 165 (not shown). Also, a single nucleotide deletion (CAAT to CA-T) was detected at position 738; this frame shift leads
to a change in amino acid sequence Regorafenib cell line after position 246. Nucleotide sequence of PCR products from several strains confirmed the deletion in manA O – Ag as characteristic of B. ovis (not shown). Table 1 Brucella strains used in this study. O-chain biosynthetic gene restriction patterns: wbk region wb region Species Biovar Serotype Strain Host/ source Geographic origin wbkE manA O-Ag manC O-Ag manB O-Ag wbkF wbkD wboA wboB manB core wa** Terrestrial mammal: B. melitensis 1 M 16 M (ATCC 23456; BCCN R1) Goat United States A A A A A A A A A A 2 A 63/9 (ATCC 23457; BCCN R2) Goat Turkey A A A B B A A A A A 3 AM Ether (ATCC 23458; BCCN R3) Goat Italy A A A B A A A A A A B. abortus 1 A 544 (ATCC 23448; BCCN R4) Cattle England A A A C A B B A A A 2 A 86/8/59 (ATCC 23449; BCCN R5) Cattle England A A A C C B B A A A 3 A Tulya (ATCC 23450; BCCN R6) Human Uganda A A A A A B B A A A 4 M 292 (ATCC 23451; BCCN R7) Cattle England A A A C A B B A A A 5 M B3196 (ATCC 23452; BCCN R8) Cattle England A A A C A B B A A A 6 A 870 (ATCC 23453; BCCN R9) Cattle Africa A A A C A B B A A A 9 M C68 (ATCC 23455; BCCN R11) Cattle England A A A C A B B A A A R 45/20 (BCCN V2) Cattle England A A A C C B B A A A B.