Genomic variations leading to alterations in cell morphology of Campylobacter spp.. Genome changes causing Campylobacter shape change

Campylobacter jejuni, the most common cause of bacterial diarrhoeal disease, is normally helical. However, it can also adopt straight rod, elongated helical and coccoid forms. Studying how helical morphology is generated, and how it switches between its different forms, is an important objective for understanding this pathogen. Here, we aimed to determine the genetic factors involved in generating the helical shape of Campylobacter. For the sequencing of M1cam helical to rod isolates, sequencing libraries were prepared using the NEBNext Ultra II DNA library prep kit (New England Biolabs). 250 ng DNA was sheared to 400 bp fragments in microTUBE screw-cap tubes in a M220 focused-ultrasonicator (Covaris). Following DNA library preparation, the library size was determined with a Bioanalyzer 2100 (Agilent), quantified using the Qubit dsDNA BR kit (Life Technologies), pooled in equal quantities, and analysed with the NEBNext library quant kit (New England Biolabs). The pooled library was subjected to 150 bp paired-end sequencing (Genomics core facility at Cancer Research UK). The read files were demultiplexed using the demuxFQ tool developed at Cancer Research UK.