Entamoeba histolytica: cloning and characterization of actin cDNA

Marion Huber, Leonard Garfinkel, Carlos Gitler, David Mirelman, Michel Revel, Shmuel Rozenblatt

Research output: Contribution to journalArticlepeer-review

Abstract

In order to study gene expression in the human parasite Entamoeba histolytica, a cDNA library of E. histolytica strain 200: NIH was constructed using the phage vector λgt10. Three cDNA clones (A,B and C) were selected for further analysis. Each of the three clones hybridized to a distinct mRNA. Two of these mRNAs were translated in vitro afer hybrid selection, and yielded distinct translation products. One of these mRNAs, selected by hybridization to clone A, encodes the most abundantly expressed protein in E. histolytica. DNA sequence analysis of this cDNA clone identified the DNA as that encoding actin. The deduced amino acid sequence of E. histolytica actin resembles both cytoplasmic and muscle actins and has unusual N-terminal glycine residue. We have shown that a family of actin genes is present in E. histolytica actin clones were obtained from a λgt10 genomic library using subcloned cDNA probes. Southern analysis of three different E. histolytica strains (200:NIH, Rhaman, and HM-1:IMSS) revealed at least four different actin genes. Strain HM-1:IMSS, however, differs by the presence of an additional actin gene.

Original languageEnglish
Pages (from-to)227-235
Number of pages9
JournalMolecular and Biochemical Parasitology
Volume24
Issue number3
DOIs
StatePublished - Jul 1987
Externally publishedYes

Keywords

  • Actin
  • DNA
  • Entamoeba histolytica
  • Multigene family
  • cDNA cloning
  • sequence

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