A HAPPY map of Cryptosporidium parvum

Abstract

We have constructed a HAPPY map of the apicomplexan parasite Crytosporidium parvum. We have placed 204 markers on the 10.4-Mb genome, giving an average marker spacing of ~50 kb, with an effective resolution of ~40 kb. HAPPY mapping (an in vitro linkage technique based on screening approximately haploid amounts of DNA by the polymerase chain reaction) is fast and accurate and is not subject to the distortions inherent in cloning, meiotic recombination, or hybrid cell formation. In addition, little genomic DNA is needed as a substrate, and the AT content of the genome is largely immaterial, making it an ideal method for mapping otherwise intractable parasite genomes. The map, covering all eight chromosomes, consists of 10 linkage groups, each of which has been chromosomally assigned. We have verified the accuracy of the map by several methods, including the construction of a > 140-kb PAC contig on chromosome VI. Less than 1% of our markers detect non-rDNA duplicated sequences.