cut-1-like genes are present in the filarial nematodes, Brugia pahangi and Brugia malayi, and, as in other nematodes, code for components of the cuticle

Mol Biochem Parasitol. 1999 Jun 25;101(1-2):173-83. doi: 10.1016/s0166-6851(99)00070-5.

Abstract

A fragment of a cut-1 like gene from the filarial nematode Brugia pahangi (designated Bp-cut-1) was isolated by PCR from genomic DNA. The sequence was used to design primers for use in RT-PCR and resulted in the isolation of a cDNA fragment from larvae in the process of the L3-L4 moult. Screening of a B. malayi genomic library identified a single clone, Bm-cut-1. Using primers designed from the Brugia sequences, semi-quantitative RT-PCR was carried out on 11 different life cycle stages chosen to cover periods around the moult and inter-moult periods. This analysis demonstrated that the cut-1 mRNA was most abundant preceding the moult, consistent with its function as a cuticular protein. Immuno-gold electron microscopy using an affinity purified antiserum raised to the highly conserved region of Ascaris CUT-1 confirmed that the protein was restricted to a tight band in the median layer of the cuticle. Despite the fact that no transcripts could be detected in mature adult worms by RT-PCR, immuno-gold microscopy revealed staining of the microfilarial cuticle within the uterus of the adult female worm, suggesting that other cut-1-like genes are present in Brugia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Brugia malayi / chemistry
  • Brugia malayi / genetics*
  • Brugia malayi / growth & development
  • Brugia pahangi / chemistry
  • Brugia pahangi / genetics*
  • Brugia pahangi / growth & development
  • Caenorhabditis elegans Proteins*
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Female
  • Gene Library
  • Genes, Helminth*
  • Helminth Proteins / analysis
  • Helminth Proteins / chemistry
  • Helminth Proteins / genetics*
  • Immunohistochemistry
  • Male
  • Microscopy, Electron
  • Molecular Sequence Data
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Caenorhabditis elegans Proteins
  • DNA, Complementary
  • Helminth Proteins
  • RNA, Messenger
  • CUT-1 protein, C elegans

Associated data

  • GENBANK/AJ012617
  • GENBANK/AJ012618
  • GENBANK/Z50044