Leishmania donovani: characterization and expression of ORFF, a gene amplified from the LDI locus

Exp Parasitol. 1999 Dec;93(4):225-30. doi: 10.1006/expr.1999.4453.

Abstract

The LD1 locus is a 27.5-kb region of chromosome 35 that is conserved among all species of Leishmania and is amplified in several different isolates. Here, we report the genomic distribution of ORFF, a gene from the LD1 region, and its expression at the RNA and protein levels in two Indian isolates of Leishmania donovani. In both of these isolates, ORFF was present as a single copy on chromosome 35. Densitometric analysis of ORFF mRNA abundance revealed relative abundance of 0.2 and 1.0 in AG83 and S-Lal, respectively. Antiserum against recombinant ORFF protein detected a protein of the predicted size ( approximately 34 kDa) in both strains. The protein is most abundant in mid-log-phase promastigotes and has a nuclear localization. The ORFF protein is preferentially expressed in L. donovani amastigotes but, in contrast, is expressed at higher levels in L. major promastigotes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Northern
  • Blotting, Southern
  • Blotting, Western
  • Cell Nucleus / chemistry
  • Densitometry
  • Gene Amplification*
  • Gene Expression Regulation*
  • Leishmania donovani / genetics*
  • Leishmania donovani / metabolism
  • Luminescent Measurements
  • Protozoan Proteins / analysis
  • Protozoan Proteins / biosynthesis
  • Protozoan Proteins / genetics*
  • RNA, Messenger / analysis
  • RNA, Protozoan / analysis

Substances

  • ORFF protein, Leishmania
  • Protozoan Proteins
  • RNA, Messenger
  • RNA, Protozoan