Campylobacter jejuni is the most commonly recognised cause of bacterial gastroenteritis in humans. Initial difficulties in culture requirements, and attempts at genetic modification held back the progress in understanding this organism compared to other enteric pathogens. However, the sequencing of 1.64 million nucleotides in the genome at the Wellcome Trust Sanger Institute¹ has heralded a new era in C. jejuni research. Using this sequence information, along with 31 predicted ORF's by TIGR, Jodi Parrish (Russell Finley's lab - Wayne State University School of Medicine) has created a C. jejuni ORF library². The proteome-scale expression clone set comprises of >1,600 ORF's. The clones were generated by efficiently utilizing the native E. coli recombination system. Recombination sites have been confirmed by sequencing, and most of the clones have been 5' sequence verified. The ORFs have a range from 45 - 4020bp in length. After expression, affinity purification can be performed on the proteins using either GST or His-tags. The clone set has many applications, including protein interactions, activity, and structural studies. Furthermore, the clones can be utilised for gene expression patterns by micro-array analysis.
Please click on Cjejuni.xls to view database. Individual clones can be ordered on GenomeCube using the Cj clone ID (e.g. Cj0748)
This is the diagrammatic representation of the expression and cloning regions of pTLJ03 ( vector map, sequence). The process of endogenous recombination in E. coli facilitates the insertion of the ORF in the correct 5' to 3' direction. The antibiotic resistance gene, encoded on the plasmid, allows for the selection of the new construct.
Source BioScience offers the complete clone set of 17 96-well microtitre plates or individual clones, for research purposes only. There is also a C. jejuni finder tool available which allows you to find the location of the clone in the 96-well plates. We can also make subsets of your chosen clones from the collection. Please contact us to discuss your requirements.
¹Parkhill J, Wren BW, Mungall K, Ketley JM, Churcher C, Basham D, Chillingworth T, Davies RM, Feltwell T, Holroyd S, Jagels K, Karlyshev AV, Moule S, Pallen MJ, Penn CW, Quail MA, Rajandream MA, Rutherford KM, van Vliet AH, Whitehead S, Barrell BG. The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences. Nature. 2000 Feb 10; 403 (6770): 665-8.
²Parrish J, Limjindaporn T, Hines JA, Liu J, Liu G, and Finley RL, Jr. High-throughput cloning of Campylobacter jejuni ORFs by in vivo recombination in Escherichia coli, Journal of Proteome Research, 2004, 3 (3), pp 582–586