Doležel, J., Vrána, J., Šafář, J., Bartoš, J., Kubaláková, M., Šimková, H.
FUNCTIONAL & INTEGRATIVE GENOMICS
12:
397-416,
2012
Klíčová slova:
Chromosome sorting, Chromosome-specific BAC libraries, Chromosome sequencing, Chromosome genomics, Genome complexity reduction, Flow cytometry, Physical mapping
Abstrakt:
Nuclear genomes of human, animals, and plants
are organized into subunits called chromosomes. When isolated
into aqueous suspension, mitotic chromosomes can be
classified using flow cytometry according to light scatter and
fluorescence parameters. Chromosomes of interest can be
purified by flow sorting if they can be resolved from other
chromosomes in a karyotype. The analysis and sorting are
carried out at rates of 102–104 chromosomes per second, and
for complex genomes such as wheat the flow sorting technology
has been ground-breaking in reducing genome complexity
for genome sequencing. The high sample rate
provides an attractive approach for karyotype analysis (flow
karyotyping) and the purification of chromosomes in large
numbers. In characterizing the chromosome complement of
an organism, the high number that can be studied using flow
cytometry allows for a statistically accurate analysis. Chromosome
sorting plays a particularly important role in the
analysis of nuclear genome structure and the analysis of
particular and aberrant chromosomes. Other attractive but
not well-explored features include the analysis of chromosomal
proteins, chromosome ultrastructure, and highresolution
mapping using FISH. Recent results demonstrate
that chromosome flow sorting can be coupled seamlessly
with DNA array and next-generation sequencing technologies
for high-throughput analyses. The main advantages are
targeting the analysis to a genome region of interest and a
significant reduction in sample complexity. As flow sorters
can also sort single copies of chromosomes, shotgun sequencing
DNA amplified from them enables the production
of haplotype-resolved genome sequences. This review
explains the principles of flow cytometric chromosome analysis
and sorting (flow cytogenetics), discusses the major uses
of this technology in genome analysis, and outlines future
directions.
Fulltext: kontaktujte autory z ÚEB
Autoři z ÚEB: Jan Bartoš,
Jaroslav Doležel,
Marie Kubaláková,
Jan Šafář,
Hana Šimková,
Jan Vrána
Čeština
English