Schistosoma mansoni: Chromosomal localization of DNA repeat elements by in situ hybridization using biotinylated DNA probes

Hirohisa Hirai, Loretta D. Spotila, Philip T Loverde

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Localization of the SMα family of repeated DNA and the rDNA repeat on the chromosomes of Schistosoma mansoni by in situ hybridization is presented. Biotinylated DNA was hybridized to target chromosomes and hybridization was detected using either alkaline phosphatase-labeled avidin or fluorescein-labeled avidin and biotinylated anti-avidin antibody. Hybridization detection using a fluorescein conjugate was more specific and sensitive with less background noise than detection with alkaline phosphatase conjugates. SMα hybridizing sequences were found dispersed throughout the genome, hybridizing to the sex chromosomes and autosomes. The SMα probe showed specific hybridization to the euchromatic gap region within the large heterochromatic block of the short arm of the W chromosome. This specific hybridization coupled with the lack of chiasma formation in this region of the ZW bivalent (presumably due to the heterochromatinization of this region) may explain the pattern of sex-specific hybridization reported for the SMα family. The rDNA repeat was localized to the secondary constriction of the short arm of chromosome 3. Specifically, the rDNA probe hybridized with the stalk of the secondary constriction and with parts of both side regions, the satellite and the short arm proper.

Original languageEnglish (US)
Pages (from-to)175-188
Number of pages14
JournalExperimental Parasitology
Volume69
Issue number1
DOIs
StatePublished - 1989
Externally publishedYes

Fingerprint

Schistosoma mansoni
DNA Probes
Ribosomal DNA
In Situ Hybridization
Avidin
Chromosomes
Constriction
Alkaline Phosphatase
DNA
Sex Chromosomes
Chromosomes, Human, Pair 3
Fluorescein
Noise
Anti-Idiotypic Antibodies
Genome

ASJC Scopus subject areas

  • Immunology
  • Parasitology
  • Infectious Diseases

Cite this

Schistosoma mansoni : Chromosomal localization of DNA repeat elements by in situ hybridization using biotinylated DNA probes. / Hirai, Hirohisa; Spotila, Loretta D.; Loverde, Philip T.

In: Experimental Parasitology, Vol. 69, No. 1, 1989, p. 175-188.

Research output: Contribution to journalArticle

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AB - Localization of the SMα family of repeated DNA and the rDNA repeat on the chromosomes of Schistosoma mansoni by in situ hybridization is presented. Biotinylated DNA was hybridized to target chromosomes and hybridization was detected using either alkaline phosphatase-labeled avidin or fluorescein-labeled avidin and biotinylated anti-avidin antibody. Hybridization detection using a fluorescein conjugate was more specific and sensitive with less background noise than detection with alkaline phosphatase conjugates. SMα hybridizing sequences were found dispersed throughout the genome, hybridizing to the sex chromosomes and autosomes. The SMα probe showed specific hybridization to the euchromatic gap region within the large heterochromatic block of the short arm of the W chromosome. This specific hybridization coupled with the lack of chiasma formation in this region of the ZW bivalent (presumably due to the heterochromatinization of this region) may explain the pattern of sex-specific hybridization reported for the SMα family. The rDNA repeat was localized to the secondary constriction of the short arm of chromosome 3. Specifically, the rDNA probe hybridized with the stalk of the secondary constriction and with parts of both side regions, the satellite and the short arm proper.

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