Using analytical ultracentrifugation to study compositional variation in vertebrate genomes.
Although much attention has recently been directed to analytical ultracentrifugation (AUC), the revival of interest has hardly addressed the applications of this technology in genome analysis, and the extent to which AUC studies can quickly and effectively complement modern sequence-based analyses of genomes, e.g. by anticipating, extending or checking results that can be obtained by cloning and sequencing. In particular, AUC yields a quick overview of the base compositional structure of a species' genome even if no DNA sequences are available and the species is unlikely to be sequenced in the near future. The link between AUC and DNA sequences dates back to 1959, when a precise linear relation was discovered between the GC (guanine+cytosine) level of DNA fragments and their buoyant density in CsCl as measured at sedimentation equilibrium. A 24-hour AUC run of a high molecular weight sample of a species' total DNA already yields the GC distribution of its genome. AUC methods based on this principle remain sensitive tools in the age of genomics, and can now be fine-tuned by comparing CsCl absorbance profiles with the corresponding sequence histograms. The CsCl profiles of vertebrates allow insight into structural and functional properties that correlate with base composition, and their changes during vertebrate evolution can be monitored by comparing CsCl profiles of different taxa. Such comparisons also allow consistency checks of phylogenetic hypotheses at different taxonomic levels. We here discuss some of the information that can be deduced from CsCl profiles, with emphasis on mammalian DNAs.
Clay
Oliver
O
Laboratory of Molecular Evolution, Stazione Zoologica Anton Dohrn, Villa Comunale, 80121 Napoli, Italy.
Douady
Christophe J
CJ
Carels
Nicolas
N
Hughes
Sandrine
S
Bucciarelli
Giuseppe
G
Bernardi
Giorgio
G
D000818:Animals; D002499:Centrifugation, Density Gradient / Q000379:methods*; D003553:Cystine / Q000032:analysis / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D004252:DNA Mutational Analysis / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D014644:Genetic Variation; D016678:Genome; D019791:Guanidine / Q000032:analysis / Q000737:chemistry; D006801:Humans; D051379:Mice; D008956:Models, Chemical; D008970:Molecular Weight; D010802:Phylogeny; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014714:Vertebrates
D003553:Cystine; D004247:DNA; D019791:Guanidine