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Occurence of subtelomeric rearrangements in the genome of the microsporidian parasite Encephalitozoon cuniculi , as revealed by a new fingerprinting procedure based on two‐dimensional pulsed field gel electrophoresis
Author(s) -
Brugère JeanFrançois,
Cornillot Emmanuel,
Méténier Guy,
Vivarès Christian P.
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(20000701)21:12<2576::aid-elps2576>3.0.co;2-d
Subject(s) - biology , pulsed field gel electrophoresis , genetics , karyotype , subtelomere , genome , amplified fragment length polymorphism , genomic dna , restriction fragment length polymorphism , chromosome , restriction fragment , microbiology and biotechnology , dna , polymerase chain reaction , gene , genotype , population , demography , sociology , genetic diversity
In Microsporidia , mitochondria‐lacking eukaryotic intracellular parasites, genomic comparisons were so far based on molecular karyotyping. The mammal‐infecting species Encephalitozoon cuniculi is characterized by a very low haploid genome size (˜2.8 Mbp) and rather high karyotype variability. Recently, we developed a two‐dimensional pulsed field gel electrophoresis (2‐D PFGE) fingerprinting technique useful for constructing a restriction map fo the genome of a mouse E. cuniculi isolate (karyotype variant A). The so‐called karyotype and restriction display 2‐D PFGE (KARD‐PFGE) protocol involved 1‐D chromosome separation, digestion with a rare cutter, Klenow radiolabeling of genomic DNA and 2‐D separation of restriction fragments followed by autoradiography. In order to assess its suitability for detecting polymorphic loci in E. cuniculi , we applied KARD‐PFGE with either Bss HII or Mlu I digestion to genome analysis of two rabbit isolates representative of two different karyotype variants (A and C). The 2‐D spot pattern of the rabbit isolate variant A is identical to the reference mouse isolate but differs greatly from the rabbit isolate variant C. Chromosomal restriction fragment length polymorphisms (RFLPs) provide strong evidence for homologous chromosomes and frequent DNA rearrangements within subtelomeric regions just upstream of the dispersed rDNA units closely associated with each chromosomal end.