Campylobacter jejuni: Methods and Protocols by James Butcher, Alain Stintzi

By James Butcher, Alain Stintzi

​ Campylobacter jejuni (C. jejuni) is frequently considered as the single of the commonest factors of bacterial gastroenteritis world wide. The objective of this quantity is to focus on key protocols for operating with C. jejuni. particularly, chapters objective to focus on contemporary advancements just about in vivo types for C. jejuni pathogenesis, various methods to isolate Campylobacter, and a structures biology method for learning the impact of all power Campylobacter gene mutants.Written within the hugely profitable Methods in Molecular Biologyseries layout, chapters contain introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, comfortably reproducible laboratory protocols, and pointers on troubleshooting and fending off recognized pitfalls.

Authoritative and state of the art, <Campylobacter jejuni: tools and Protocols encourages latest Campylobacter researchers to hire novel the right way to additional their very own learn and in addition encourages new researchers to incorporate Campylobacter of their destiny examine initiatives.

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Extra resources for Campylobacter jejuni: Methods and Protocols

Example text

C. jejuni colonizes the intestinal tract of poultry to high numbers, and regardless of the biosecurity measures aiming at reducing C. jejuni in the primary production, poultry meat remains to be the major source of the disease [1]. Since bacteriophages (phages) are often found in niches associated with their hosts, a number of samples have been used for phage isolation such as poultry meat [2], poultry feces [3–5] and intestines [6, 7], pig manure, slaughterhouse wastewater, and sewage [8, 9]. Here, we describe modifications of general techniques, which are needed to successfully isolate C.

We commonly employ a single-cell lysis buffer (SCLB) [20] as substitute for DI water to prepare less crude DNA extract, which appears to yield more consistent and reliable PCR results. Methods to Study Antimicrobial Resistance in Campylobacter jejuni 39 As with the boiling method, a few colonies are suspended in 40 μL SCLB, which is then heated to 80 °C for 10 min and cooled down to 55 °C for 10 min in a thermocycler to lyse bacterial cells. The suspension is then diluted 1:2 in DI water and centrifuged at 4500 × g for 30 s to remove cellular debris.

Therefore, ribosomal binding sites (RBS) and start codons are key target sites for antisense inhibition by PNA. Several studies have shown that the start codon region is susceptible to antisense inhibition [7–9], and the RBS is also an useful target in C. jejuni [6]. However, a golden rule does not seem to exist yet in PNA design to pinpoint most the effective inhibition site. Based on our previous study to inhibit the expression of CmeABC, a multidrug efflux pump, the choice of targeting flanking regions of the RBS or the start codon also substantially affect the efficacy of antisense inhibition by PNA [6].

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