Open AccessGenotyping of Ochrobactrum anthropi by recA ‐based comparative sequence, PCR‐RFLP, and 16S rRNA gene analysis
Author(s) -
Scholz Holger C.,
Tomaso Herbert,
Dahouk Sascha Al,
Witte Angela,
Schloter Michael,
Kämpfer Peter,
Falsen Enevold,
Neubauer Heinrich
Publication year - 2006
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00153.x
Subject(s) - haeiii , biology , genetics , phylogenetic tree , restriction fragment length polymorphism , brucella , 16s ribosomal rna , sequence analysis , genotyping , microbiology and biotechnology , polymerase chain reaction , gene , genotype , virology , brucellosis
A recA ‐PCR restriction fragment length polymorphism assay was developed to study intraspecies variation among Ochrobactrum anthropi . Primers deduced from the known recA gene sequence of the genetically closely related genus Brucella allowed the specific amplification of a 1065 bp recA fragment from each of the 38 O. anthropi and the eight Brucella strains investigated. RecA was also amplified from the type strains of O. intermedium, O. tritici , and O. lupini but could not be generated from O. grignonense and O. gallinifaecis . Subsequent comparative recA sequence‐ and Hae III‐ recA restriction fragment length polymorphism analysis identified nine different genospecies among the tested 38 O. anthropi isolates, whereas the recA sequences of the Brucella spp. were indistinguishable. Furthermore, Brucella spp., O. anthropi, O. intermedium , and O. tritici were clearly separated from each other by means of their recA sequences and Hae III restriction patterns. Five strains of uncertain species status listed in the Culture Collection University of Göteborg bacterial culture collection as O. anthropi were characterized by recA analysis, and their phylogenetic position within the Brucella–Ochrobactrum group was determined. In summary, recA ‐sequence analysis provides a new reliable molecular subtyping tool to study the phylogeny of the Ochrobactrum taxon at both the inter‐ and intraspecies level.