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

Kami Kim, Lisa Goozé, Carolyn Petersen, Jiri Gut, Richard G. Nelson

Research output: Contribution to journalArticle

77 Scopus citations

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 β-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 γ-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.

Original languageEnglish (US)
Pages (from-to)105-113
Number of pages9
JournalMolecular and Biochemical Parasitology
Volume50
Issue number1
DOIs
Publication statusPublished - Jan 1992

    Fingerprint

Keywords

  • Actin
  • Apicomplexa
  • Cryptosporidium parvum
  • Gliding motility
  • Molecular karyotype
  • Pulsed field gel electrophoresis

ASJC Scopus subject areas

  • Parasitology
  • Molecular Biology

Cite this