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Related Experiment Videos

Female-specific sequences isolated from Schistosoma mansoni by representational difference analysis

A C Drew1, P J Brindley

  • 1Molecular Parasitology Unit, Queensland Institute of Medical Research, Royal Brisbane Hospital, Brisbane, Australia.

Molecular and Biochemical Parasitology
|May 1, 1995
PubMed
Summary

Representational difference analysis (RDA) identified two new female-specific repetitive DNA sequences in Schistosoma mansoni. These sequences, SM alpha fem-1 and W2, are located on the W chromosome.

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Area of Science:

  • Genetics and Genomics
  • Molecular Biology
  • Parasitology

Background:

  • Understanding genetic differences between sexes is crucial for studying reproduction and evolution in parasitic organisms.
  • Schistosoma mansoni, a significant human parasite, presents challenges in sex determination research due to limited knowledge of sex-specific genetic elements.

Purpose of the Study:

  • To isolate and characterize female-specific DNA sequences in Schistosoma mansoni using a molecular approach.
  • To identify novel genetic elements potentially involved in sex determination or differentiation within the S. mansoni genome.

Main Methods:

  • Utilized Representational Difference Analysis (RDA), a PCR-based modification of subtractive hybridization, to enrich for female-specific sequences.
  • Employed HindIII-derived amplicons and an excess of male DNA to subtract common sequences from female genomic DNA.

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  • Performed three rounds of RDA to isolate and identify distinct female-specific repetitive elements.

    • Main Results:

    • Successfully isolated two distinct female-specific repetitive DNA sequences from the Schistosoma mansoni genome.
    • One sequence, SM alpha fem-1, showed 76% homology to the SM alpha retroposon family, representing a W chromosome-specific variant.
    • The second sequence, W2, is a novel female-specific repetitive sequence, with both elements appearing as tandem repeats on the W chromosome.

    Conclusions:

    • Representational Difference Analysis is an effective method for isolating sex-specific sequences in Schistosoma mansoni.
    • The identified sequences, SM alpha fem-1 and W2, are novel W chromosome-specific elements, providing insights into the parasite's sex determination.
    • RDA holds potential for defining differentiating traits and sequences across various schistosome species.