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Isolation, sequence and molecular karyotype analysis of the actin gene of Cryptosporidium parvum.

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Affiliations

  • 1. Parasitology Laboratory, San Francisco General Hospital, CA.
      Authors
    • Kim K 1
    (1 author)

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Molecular and Biochemical Parasitology, 01 Jan 1992, 50(1):105-113
https://doi.org/10.1016/0166-6851(92)90248-i PMID: 1542305 

Abstract 

Actin is an ubiquitous and highly conserved microfilament protein which is hypothesized to play a mechanical, force-generating role in the unusual gliding motility of sporozoan zoites and in their active penetration of host cells. We have identified and isolated an actin gene from a Cryptosporidium parvum genomic DNA library using a chicken beta-actin cDNA as an hybridization probe. The nucleotide sequences of two overlapping recombinant clones were identical and the amino acid sequence deduced from the single open reading frame was 85 % identical to the P. falciparum actin I and human gamma-actin proteins. The predicted 42 106-Da Cryptosporidium actin contains 376 amino acids and is encoded by a single-copy gene which contains no introns. The nucleic acid coding sequence is 72% biased to the use of A or T in the third position of codons. Chromosome-sized DNA released from intact C. parvum oocysts was resolved by OFAGE into 5 discrete ethidium bromide-staining DNAs ranging in size from 900 to 1400 kb; the cloned C. parvum actin gene hybridized to a single chromosomal DNA of approximately 1200 kb.

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