DNA reassociation kinetics
Answers
Explanation:
DNA reassociation kinetics were studied, by means of the hydroxyapatite chromatography method, for four species of Amphibians with different nuclear DNA content: Xenopus laevis (3 pg DNA per haploid genome) and Bufo bufo (7 pg) of the Anura subclass and Trituras cristatus (23 pg) and Necturus maculosus (52 pg) of the Urodela subclass.
Principle behind Cot curve: The rate at which a particular sequence of a DNA reassociates(renatures) is proportional to the number of times it is found in the DNA molecule. That means, Rate of Renaturation of a sequence of a DNA molecule ∝ number of times the sequence is present in the DNA molecule.
Within each subclass the two species studied were found to have about the same absolute amount of unique DNA. The differences of total nuclear DNA can be accounted for by quantitative variations of the repetitive sequence classes, at least in part due to changes in the number of copies of the various sequences. On the contrary the great difference in nuclear DNA between the two subclasses, Anura and Urodela, involves all sequence classes in parallel; the slowly reassociating fraction appears to be unique in spite of a tenfold difference in absolute amount.
DNA reassociation kinetics have been partly elucidated for the higher crabs C. borealis and L. emarginata, using calf thymus DNA as a standard. These crabs contain no detectable repeated DNA in the approximate multiplicity frequency range 2-100 copies, which is unusual for invertebrate DNAs. Each species contains a component renaturing at an intermediate rate, and also a very rapidly renaturing fraction. The very rapidly renaturing fraction is considerably larger than the cesium chloride-resolvable satellites of each species. The fraction reassociating at an intermediate rate includes sequences with a reiteration frequency of up to 9.0 X 10(4) copies. This is unusually high for invertebrate DNAs. The nearly exact correlation between kinetic complexity and independently determined haploid genome size leads to the conclusion that the most slowly renaturing sequences of both crab species are present only once per haploid genome. Therefore the chromatids of these species are uninemic structures, and there has been no detectable occurrence of polyploid speciation in the recent evolutionary history of either species.
The dependence of reassociation kinetics on DNA fragment length for the four species indicates for all of them an interspersed organization of the various sequence classes