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Abstract
Fluorescence in situ hybridization (FISH) allows identification of particular chromosomes and their rearrangements. Using FISH with signal enhancement via antibody amplification and enzymatically catalysed reporter deposition, we evaluated applicability of universal cytogenetic markers, namely 18S and 5S rDNA genes, U1 and U2 snRNA genes, and histone H3 genes, in the study of the karyotype evolution in moths and butterflies. Major rDNA underwent rather erratic evolution, which does not always reflect chromosomal changes. In contrast, the hybridization pattern of histone H3 genes was well conserved, reflecting the stable organisation of lepidopteran genomes. Unlike 5S rDNA and U1 and U2 snRNA genes which we failed to detect, except for 5S rDNA in a few representatives of early diverging lepidopteran lineages. To explain the negative FISH results, we used quantitative PCR and Southern hybridization to estimate the copy number and organization of the studied genes in selected species. The results suggested that their detection was hampered by long spacers between the genes and/or their scattered distribution. Our results question homology of 5S rDNA and U1 and U2 snRNA loci in comparative studies. We recommend the use of histone H3 in studies of karyotype evolution.
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1 University of South Bohemia, Faculty of Science, České Budějovice, Czech Republic (GRID:grid.14509.39) (ISNI:0000 0001 2166 4904); Biology Centre CAS, Institute of Entomology, České Budějovice, Czech Republic (GRID:grid.418338.5) (ISNI:0000 0001 2255 8513); European Molecular Biology Laboratory, Heidelberg, Germany (GRID:grid.4709.a) (ISNI:0000 0004 0495 846X)
2 University of South Bohemia, Faculty of Science, České Budějovice, Czech Republic (GRID:grid.14509.39) (ISNI:0000 0001 2166 4904); Biology Centre CAS, Institute of Entomology, České Budějovice, Czech Republic (GRID:grid.418338.5) (ISNI:0000 0001 2255 8513)
3 University of South Bohemia, Faculty of Science, České Budějovice, Czech Republic (GRID:grid.14509.39) (ISNI:0000 0001 2166 4904); Biology Centre CAS, Institute of Entomology, České Budějovice, Czech Republic (GRID:grid.418338.5) (ISNI:0000 0001 2255 8513); University of Groningen, Groningen Institute for Evolutionary Life Sciences, Groningen, The Netherlands (GRID:grid.4830.f) (ISNI:0000 0004 0407 1981)
4 The Pirbright Institute, Surrey, UK (GRID:grid.63622.33) (ISNI:0000 0004 0388 7540)
5 Biology Centre CAS, Institute of Entomology, České Budějovice, Czech Republic (GRID:grid.418338.5) (ISNI:0000 0001 2255 8513)