Forward Genetic Analysis of the Apicomplexan Cell Division Cycle in Toxoplasma gondii

摘要: Apicomplexa are obligate intracellular pathogens that have fine-tuned their proliferative strategies to match a large variety of host cells. A critical aspect this adaptation is flexible cell cycle remains poorly understood at the mechanistic level. Here we describe forward genetic dissection apicomplexan using Toxoplasma model. By high-throughput screening, isolated 165 temperature sensitive parasite growth mutants. Phenotypic analysis these mutants suggests regulated progression through with defined phases and checkpoints. These analyses also highlight importance peculiar intranuclear spindle as physical hub regulation. To link phenotypes genes, developed robust complementation system based on genomic cosmid library. Using approach, so far complemented 22 identified 18 candidate loci, eight which were independently confirmed set sequenced arrayed cosmids. For three loci mutant allele. The genes include regulators formation, nuclear trafficking, protein degradation. approach described here should be widely applicable numerous essential aspects biology.