Genetic relationships amongPseudomonas stutzeristrains based on molecular typing methods

Abstract

A . B EN N AS AR , C. GU A SP , M . TE SA R AN D J . LA LU C AT . 1998. Detailed characterization of the genetic variability among strains belonging to Pseudomonas stutzeri was achieved using different rapid molecular typing methods based on polymerase chain reaction (PCR), Southern blot and Western blot. Consensus motifs complementary to fragments of repetitive elements dispersed throughout the genomes of bacteria were used as primers and allowed differentiation at subspecies levels. Further and simple differentiation was also achieved based on the direct amplification of spacer regions between 16S and 23S rRNA, combined with single-strand conformation polymorphism (SSCP) analysis of the generated fragments. These methods are fast, sensitive, reliable for determining relationships, and have demonstrated a great genetic diversity among the strains of Ps. stutzeri studied in agreement with the heterogeneous phenotypic traits of the species.

The potential for intraspecific horizontal gene exchange by natural genetic transformation: sexual isolation among genomovars of Pseudomonas stutzeri The EMBL accession numbers for the sequences reported in this paper are given in Methods.

“…The genomovar structure of P. stutzeri is consistent with the finding of seven genotypic groups in a genomic fingerprint analysis (Sikorski et al, 1999). Variable grouping of various strains in cluster analyses of genomic fingerprints and macrorestriction patterns indicated that chromosomal rearrangements, presumably also a result of horizontal gene transfer, have occurred and contributed to the extremely high genotypic diversity within this species (Bennasar et al, 1998 ;Ginard et al, 1997 ;Rainey et al, 1994 ;Sikorski et al, 1999).…”

Biology of Pseudomonas stutzeri

“…Similar genomic relationships have been revealed by PCR amplification of several genes (16S rRNA, internal transcribed spacer region 1 [ITS1], ITS2, and rpoB) and by analyzing the RFLPs generated by several restriction enzymes (25,133,325). These methods have confirmed the high genetic diversity of the species, the consistency of genomic groups (genomovars), and the usefulness of the patterns generated for strain identification.…”

“…However, a similar procedure, based on Western blotting and immunological fingerprinting of whole-cell proteins using the polyclonal antibody Ab160, raised against Pseudomonas fluorescens MT5-called Westprinting (360)-produced a typical protein profile for each strain. Computer-assisted comparisons revealed a distribution in groups that agreed with the strains' genomovar distribution at different similarity levels (25).…”

“…Bennasar et al (25) revealed genetic diversity and the relationships among P. stutzeri strains by rapid molecular typing methods. Repetitive extragenic palindromic PCR and enterobacterial repetitive intergenic consensus PCR analyses, based on DNA consensus sequences, generated fingerprints that were then computer analyzed.…”

Inv A gene specific PCR for detection of Salmonella from broilers

“…and biochemical method and then they were Established conventional methods to detect and confirmed by invA specific PCR methods. identify Salmonella are time consuming and include Materials and Methods selective enrichment and plating followed by biochemical tests (Bennasar et al, 2000;Burtscher et al, Samples from broiler carcasses and culture 1999; Chiu and Jonathan, 1996). Several techniques methods: Sixty samples of broilers were collected have been developed for the improvement in the from slaughter houses of Namakkal.…”