The PlasmoGEM vector resource and its use in high-throughput genetic screening

Bushell E, Gomes AR, Sanderson T, Anar B, Girling G, Herd C, Metcalf T, Modrzynska K, Schwach F, Martin RE, Mather MW, McFadden GI, Parts L, Rutledge GG, Vaidya AB, Wengelnik K, Rayner JC, Billker O: Functional Profiling of a Plasmodium Genome Reveals an Abundance of Essential Genes. Cell(170), 260-272, DOI

Gomes AR, Bushell E, Schwach F, Girling G, Anar B, Quail M, Herd C, Pfander C, Modrzynska K, Rayner JC and Billker O: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite. Cell Host & Microbe(17), 404-413, DOI:

Description of the PlasmoGEM software and web portal

Schwach F, Bushell E, Gomes AR, Anar B, Girling G, Herd C, Rayner JC and Billker O: PlasmoGEM, a database supporting a community resource for large-scale experimental genetics in malaria parasites. Nucleic Acids Research(43), D1176-82, DOI:

First publication describing methodology and technology adaptation to P. berghei

Pfander C, Anar B, Schwach F, Otto TD, Brochet M, Volkmann K, Quail MA, Pain A, Rosen B, Skarnes WC, Rayner JC and Billker O: A scalable pipeline for highly effective genetic modification of a malaria parasite. Nature Methods Volume 8, 1078-1082, DOI:

Book chapters describing methodology for vector production in more detail

Pfander C., Bronner-Anar B., Girling G., Bushell E. S. C. , Schwach F., Rayner J. C., and Billker O. (2013) Constructing Plasmodium berghei gene targeting vectors using recombineering. Methods in Malaria Research, 6th Edition

The N15-phage linear vector system (pJAZZ, Lucigen) used to generate P. berghei genomic library

Ravin, N. V., Kuprianov, V. V., Gilcrease, E. B. & Casjens, S. R. (2003) Bidirectional replication from an internal ori site of the linear N15 plasmid prophage. NAR 31, 6552-6560. Godiska R, Mead D, Dhodda V, Wu C, Hochstein R, Karsi A, Usdin K, Entezam A, Ravin N. (2010) Linear plasmid vector for cloning of repetitive or unstable sequences in Escherichia coli. NAR 38, e88.

Recombineering principles and methodology

Zhang, Y., Buchholz, F., Muyrers, J. P. & Stewart, A. F. (1998) A new logic for DNA engineering using recombination in Escherichia coli. Nature Genetics 20, 123-128. DOI: Wang, J. et al. (2006) An improved recombineering approach by adding RecA to lambda Red recombination. Mol. Biotech. 32, 43-53. PMID: 16382181

P. berghei positive and negative selectable marker system

Braks JA, Franke-Fayard B, Kroeze H, Janse CJ, Waters AP (2006) Development and application of a positive-negative selectable marker system for use in reverse genetics in Plasmodium. Nucleic Acids Res: 34 (5):e39.