Showing papers by "John C. Avise published in 1981"

The use of restriction endonucleases to measure mitochondrial DNA sequence relatedness in natural populations. III. Techniques and potential applications.

Abstract: Restriction endonucleases and agarose gel electrophoresis have been used to demonstrate extensive nucleotide sequence diversity in mitochondrial DNA (mtDNA) within and between conspecific populations of rodents and other mammals. Cleavage of mtDNA samples with a relatively small number of endonucleases provides information concerning the phylogenetic relatedness of individual organisms which cannot now be readily obtained by any other type of molecular analysis. This information is qualitatively different from that available from the study of nuclear genes or gene products because the mitochondrial genome is inherited intact from the female parent and is not altered by recombination or meiotic segregation. The requirements for large tissue samples and laborious DNA purification procedures have imposed severe limitations on the kinds of population surveys in which this technique could be utilized. Here, we show that these difficulties can be overcome by using DNA-DNA hybridization to detect minute amounts of mtDNA in crude tissue fractions which can be more easily and rapidly prepared from very small amounts of tissue without the use of expensive and immobile laboratory equipment. The techniques are described in detail in an effort to make restriction analysis of mtDNA available to biologists who may be unfamiliar with current DNA technology.

Genetic divergence between rodent species assessed by using two-dimensional electrophoresis

Abstract: O'Farrell's technique of two-dimensional gel electrophoresis (2-DGE) has previously been applied to the study of intrapopulation genetic variation. This approach assays a larger, and in part nonoverlapping, cohort of protein encoding loci compared to conventional one-dimensional electrophoretic procedures (SGE) and has revealed substantially lower levels of mean heterozygosity. Here we extend this approach to analyze levels and patterns of genetic differentiation between species. We have used 2-DGE to compare an average of 189 polypeptides between six species of wild mice representing levels of evolutionary divergence ranging from different subspecies to different families. The magnitude of protein divergence estimated by 2-DGE was on the average only about one-half that predicted by SGE. This discrepancy may result from differences in sensitivities between the techniques or differences in the mean level of variation and divergence between the sets of loci assayed by the two methods. Nonetheless, the ranking of genetic distances by 2-DGE was identical to that by SGE. Thus, these results support the use of the simpler SGE techniques to estimate relative levels of genetic divergence.