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Infection and Immunity, August 1999, p. 3960-3969, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
A Random Survey of the Cryptosporidium
parvum Genome
Chang
Liu,
Vladimir
Vigdorovich,
Vivek
Kapur, and
Mitchell S.
Abrahamsen*
Department of Veterinary PathoBiology,
University of Minnesota, St. Paul, Minnesota
Received 29 January 1999/Returned for modification 31 March
1999/Accepted 25 May 1999
Cryptosporidium parvum is an obligate intracellular
pathogen responsible for widespread infections in humans and animals. The inability to obtain purified samples of this organism's various developmental stages has limited the understanding of the biochemical mechanisms important for C. parvum development or
host-parasite interaction. To identify C. parvum genes
independent of their developmental expression, a random
sequence analysis of the 10.4-megabase genome of C. parvum was undertaken. Total genomic DNA was sheared by
nebulization, and fragments between 800 and 1,500 bp were gel purified
and cloned into a plasmid vector. A total of 442 clones were
randomly selected and subjected to automated sequencing by using one or
two primers flanking the cloning site. In this way, 654 genomic survey sequences (GSSs) were generated, corresponding to
>320 kb of genomic sequence. These sequences were assembled into 408 contigs containing >250 kb of unique sequence, representing ~2.5% of the C. parvum genome. Comparison of the GSSs
with sequences in the public DNA and protein databases revealed that
107 contigs (26%) displayed similarity to previously identified
proteins and rRNA and tRNA genes. These included putative genes
involved in the glycolytic pathway, DNA, RNA, and protein metabolism,
and signal transduction pathways. The repetitive sequence elements identified included a telomere-like sequence containing hexamer repeats, 57 microsatellite-like elements composed of dinucleotide or
trinucleotide repeats, and a direct repeat sequence. This study demonstrates that large-scale genomic sequencing is an
efficient approach to analyze the organizational characteristics and
information content of the C. parvum genome.
*
Corresponding author. Mailing address: Department of
Veterinary PathoBiology, University of Minnesota, 1988 Fitch
Ave., St. Paul, MN 55108. Phone: (612) 624-1244. Fax: (612)
625-0204. E-mail: abrah025{at}tc.umn.edu.
Infection and Immunity, August 1999, p. 3960-3969, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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